Objective To explore the mechanism of apoptosis after cryotherapy on tumors.Methods (1) GL261 glioma cells (1×107 cell/10 μL) were injected into the subcutaneous one ofC57 mice to establish tumor-bearing mouse models;when the diameter of tumor reached to 15-20 mm,the mice were randomly divided into cryogenic treatment group and sham-operated group (n=1 0);mice in the cryogenic treatment group were given surgical cryotherapy,while those in the sham-operated group only performed surgery without cryotherapy.TUNEL was used to detect the cell apoptosis in glioma tissues 12 and 24 h after operation;and Western blotting was employed to detect the protein expressions of pro-caspase-8,pro-caspase-9 and poly-ADP-ribose polymerase (PARP).(2) GL261 glioma cells were divided into control group and one time cryogenic release group,and DMEM and one time of cryogenic release were given to the two groups,respectively;12 h after the treatment,TUNEL was used to observe the cell apoptosis in glioma tissues,and Western blotting was employed to detect the protein expressions.Results (1) TUNEL indicated that the cells in the S1 region of the glioma tissues from mice in the cryogenic treatment group were uniformly died;significant apoptosis was noted in cells of the S2 region at 12 h after treatment;while,24 h after treatment,S1 region still showed uniform necrosis,S2 region showed apoptotic regression,and S3 region showed new apoptosis at the target side.Western blotting indicated that pro-caspase-9 and PARP protein expressions at the S2 region were signficantly decreased as compared with those at the S1,S3 and S4 regions (P＜0.05),and pro-caspase-8 protein expression at the S3 region were signficantly reduced as compared with those at the S1,S2 and S4 regions in the cryogenic treatment group (P＜0.05).(2) TUNEL showed that significantly increased GL261 glioma cell apoptosis rate was noted in the one time cryogenic release group as compared with that in the control group (P＜0.05);Western blotting indicated that as compared with the control group,the cryogenic release group had significantly decreased pro-caspase-8 and pro-caspase-9 expressions (P＜0.05).Conclusion Cryogenic release or substances released from tumor tissues after cryoablation shows an effect on promoting apoptosis.

Objective:To explore the microcirculation formation mechanism of contrast-enhanced ultrasonography imaging performance in rabbits with limb muscle crush injury.Methods:Seventy-two New Zealand white rabbits were randomly divided into two groups. A limb muscle crush injury model was created by airing a balloon cuff device with a force of 40 kPa. Contrast-enhanced ultrasonography parameters were detected in the first group.In vivo microcirculation parameters were detected in the second group. Fine blood vessel diameter and blood flow velocity were calculated before extrusion and 0.5 h, 2 h, 6 h, 24 h, and 3 days after decompression.Results:Totally six animals died before the end of the experiment (3 rabbits in ultrasonic imaging and microcirculation detection groups, respectively). Compared with the uninjured muscle, the reperfusion of the injured muscle showed early and high enhancement in contrast-enhanced ultrasonography images. The peak intensity and area under the curve were significantly higher than those of the control subgroup at each time point after decompression(all P<0.05), and reached the peak at 24 h after decompression. The time-intensity curve showed a trend of rapid elevation and gradual drop. In the microcirculation group, compared with the control group, fine artery and vein diameters in the experimental group were wider and the blood flow velocity was slower, especially in the fine veins(all P<0.05). At each time point after decompression, there was a statistical difference between the control subgroup and the control subgroup, and the change reached the peak at 24 h after decompression. The blood flow state showed that the arterioles were dominated by linear flow and linear grain flow at each time point after decompression, and linear grain flow, grain flow and grain pendulum flow were observed in the fine veins. Blood flow stagnation and adhesion of white blood cells and white microthrombus were also observed in the fine veins. Correlation analysis showed that the peak intensity and area under the curve were positively correlated with the inner diameter of arterioles and veins( r=0.84, 0.94; r=0.85, 0.94; all P<0.05), and negatively correlated with the flow velocity of arterioles and veins( r=-0.94, -0.96; r=-0.93, -0.96; all P<0.05). Conclusions:In vivo microcirculation detection can reflect changes in muscle microvascular diameter and blood flow velocity, which have a correlation with quantitative ultrasound imaging parameters.

β-glucosidase has important applications in food, medicine, biomass conversion and other fields. Therefore, exploring β-glucosidase with strong stability and excellent properties is a research hotspot. In this study, a GH3 family β-glucosidase gene named Iubgl3 was successfully cloned from Infirmifilum uzonense. Sequence analysis showed that the full length of Iubgl3 was 2 106 bp, encoding 702 amino acids, with a theoretical molecular weight of 77.0 kDa. The gene was cloned and expressed in E. coli and the enzymatic properties of purified IuBgl3 were studied. The results showed that the optimal pH and temperature for pNPG hydrolysis were 5.0 and 85 ℃, respectively. The enzyme has good thermal stability, and more than 85% of enzyme activity can be retained after being treated at 80 ℃ for2 h. This enzyme has good pH stability and more than 85% of its activity can be retained after being treated at pH 4.0-11.0 for 1 h. It was found that the enzyme had high hydrolysis ability to p-nitrophenyl β-d-glucoside (pNPG) and p-nitrophenyl β-d-xylopyranoside (pNPX). When pNPG was used as the substrate, the kinetic parameters K_m and V_max were 0.38 mmol and 248.55 μmol/(mg·min), respectively, and the catalytic efficiency k_cat/K_m was 6 149.20 s^-1mmol^-1. Most metal ions had no significant effect on the enzyme activity of IuBgl3. SDS completely inactivated the enzyme, while EDTA increased the enzyme activity by 30%. This study expanded the β-glucosidase gene diversity of the thermophilic archaea GH3 family and obtained a thermostable acid bifunctional enzyme with good industrial application potential.
beta-Glucosidase/chemistry* ; Archaea/metabolism* ; Escherichia coli/metabolism* ; Hydrogen-Ion Concentration ; Temperature ; Glucosides ; Enzyme Stability ; Substrate Specificity ; Kinetics

Objective:To compare the efficacy of femoral neck system (FNS) and cannulated compression screw (CCS) in the treatment of femoral neck fracture in the young and middle-aged patients.Methods:A retrospective cohort study was conducted to analyze the clinical data of 91 patients with femoral neck fracture admitted to Fuzhou Second Hospital from July 2020 to December 2021, including 52 males and 39 females; aged 23-65 years [(48.9±10.3)years]. Garden classification of the fracture found that 31 patients were with type I, 9 with type II, 12 with type III and 39 with type IV. Pauwels classification of the fracture found that 7 patients were with type I, 33 with type II and 51 with type III. A total of 53 patients were treated with FNS fixation (FNS group) and 38 patients with CCS fixation (CCS group). The operation time, intraoperative blood loss, Haidukewych fracture reduction quality, hospitalization time, Barthel index, fracture healing time, and weight-bearing time were compared between the two groups. The hip function was assessed by Harris hip score in both groups at postoperative 3 months, 6 months and 1 year and at the final follow-up. The incidences of postoperative complication and secondary surgery were also compared between the two groups.Results:All the patients were followed up for 15-31 months [(22.2±5.5)months]. There were no significant differences in the operation time, Haidukewych fracture reduction quality, hospitalization time, or Barthel index (all P>0.05). The intraoperative blood loss in the FNS group was 50.0(20.0,85.0)ml, which was more than that in the CCS group [20.0(10.0,50.0)ml] ( P<0.01). The fracture healing time, partial weight-bearing time, and full weight-bearing time in the FNS group [4.0(3.0,5.0)months, 3.0(2.0,3.0)months, 5.0(4.5,6.0)months] were shorter than those in the CCS group [6.0(5.0,7.0)months, 3.0(2.8,4.0)months, 6.0(6.0,7.0)months] (all P<0.01). The Harris hip score at postoperative 3 months, 6 months and 1 year and at the final follow-up were 74.0(71.0,77.0)points, 87.0(84.0,88.5)points, 91.0(88.0,95.0)points, and 94.0(91.0,96.0)points in the FNS group, significantly higher than those in the CCS group [73.0(70.0,74.0)points, 82.5(79.8,87.0)points, 88.0(83.5,91.0)points, 89.0(84.0,93.0)points] (all P<0.05 or 0.01). There were no statistically significant differences in the incidences of postoperative complication or secondary surgery between the two groups (all P>0.05). Conclusion:Compared with CCS, FNS can shorten fracture healing time, allow patients to carry full weight as soon as possible, and significantly improve hip function in the treatment of middle-aged and young adults with femoral neck fracture, although there is more intraoperative blood loss.

Objective To measure the parameters of the specific electrode trajectory angles under the condition that the trajectory length in the nuclei could be as long as possible by using 3D reconstruction of subthalamic nucleus (STN).Methods A 3D atlas of deep brain nuclei was built on the 3D reconstruction station,and the parameters of STN were measured.MR imaging of 30 Parkinson's disease (PD) patients was matched to the 3D atlas to acquire the average nucleus information.Results The STN spatial location parameters showed no significant statistical difference with imaging of these 30 patients (95％ confidence interval of difference value＜l),indicating that this 3D atlas enjoyed good representative value.After simulation,the optimal trajectory angles were achieved:the combination angles ofring/slide:55°-57°/17°-19°,which extended the trajectory within the nucleus to about (5.7±0.3)mm.Conclusion The 3D atlas shows high representative of locating deep brain nucleus and could be registrated well to particular patient's images.After simulation on the atlas,the optimal trajectory angles parameters can be obtained,which shows the specific operation of achieving longer length of electrode in nucleus.

Objective:To investigate the incidence of and risk factors for femoral neck shortening in young and middle-aged patients with displaced femoral neck fracture treated by a femoral neck system (FNS).Methods:A retrospective study was conducted to analyze the clinical data of 135 patients with displaced femoral neck fracture who had been treated by closed reduction and FNS internal fixation at Department of Orthopaedics, The Second Hospital of Fuzhou from December 2019 to December 2021. There were 85 males and 50 females, with an age of 51 (42, 57) years and a body mass index of 22.6 (20.9, 24.2) kg/m 2. According to the Garden classification, there were 29 cases of type Ⅲ and 106 cases of type Ⅳ. Femoral neck shortening was observed. The patients' gender, age, side of shortening, body mass index, fracture type, high-energy injury, time from injury to surgery, cortical comminution at the fracture end, reduction quality (Garden index), partial weight bearing time, complete weight bearing time, and functional evaluation (Harris score) were recorded for a univariate analysis. Multivariate logistic regression was used to analyze the influencing factors for moderate to severe shortening. Results:All the 135 patients were followed up for 17.2 (15.0, 20.2) months. Different degrees of femoral neck shortening happened in all patients, yielding an incidence of 100%(135/135). The femoral neck shortening was 4.2 (2.6, 7.7) mm in displaced femoral neck fractures fixated by FNS. The shortening continued to worsen within the first 3 months after operation, reaching a stable state in the first 6 months. Persistent shortening hardly occurred after 6 months. According to the shortening distance, the patients were divided into a mild shortening group (76 cases <5 mm) and a moderate to severe shortening group (59 cases ≥ 5 mm). The Harris score at the last follow-up in the mild shortening group (91.7 ± 2.3) was significantly higher than that in the moderate to severe shortening group (87.9 ± 3.8) ( P<0.001). The fracture healing rate was 98.7% (75/76) in the mild shortening group and 98.3% (58/59) in the moderate to severe shortening group, showing no significant difference ( P=1.000). Logistic regression analysis found that the risk factors for moderate to severe shortening were fracture type (type Ⅳ) ( OR = 10.227, 95% CI: 1.845 to 56.693, P = 0.008), cortical comminution at the fracture end (if present) ( OR=52.697, 95% CI: 9.870 to 281.352, P<0.001), and partial weight bearing time <4 weeks ( OR = 0.168, 95% CI: 0.032 to 0.893, P = 0.036). Conclusions:The incidence of femoral neck shortening may be high in young and middle-aged patients with displaced femoral neck fracture treated by FNS, leading to decreased hip function. The main risk factors for femoral neck shortening are fracture type (Garden type Ⅳ), cortical comminution at the fracture end and partial weight-bearing time less than 4 weeks.

β-glucosidase has important applications in food, pharmaceutics, biomass conversion and other fields, exploring β-glucosidase with strong adaptability and excellent properties thus has received extensive interest. In this study, a novel glucosidase from the GH1 family derived from Cuniculiplasma divulgatum was cloned, expressed, and characterized, aiming to find a better β-glucosidase. The amino acid sequences of GH1 family glucosidase derived from C. divulgatum were obtained from the NCBI database, and a recombinant plasmid pET-30a(+)-CdBglA was constructed. The recombinant protein was induced to express in Escherichia coli BL21(DE3). The enzymatic properties of the purified CdBglA were studied. The molecular weight of the recombinant CdBglA was 56.0 kDa. The optimum pH and temperature were 5.5 and 55 ℃, respectively. The enzyme showed good pH stability, 92.33% of the initial activity could be retained when treated under pH 5.5-11.0 for 1 h. When pNPG was used as a substrate, the kinetic parameters K_m, V_max and K_cat/K_m were 0.81 mmol, 291.99 μmol/(mg·min), and 387.50 s^-1 mmol^-1, respectively. 90.33% of the initial enzyme activity could be retained when CdBglA was placed with various heavy metal ions at a final concentration of 5 mmol/L. The enzyme activity was increased by 28.67% under 15% ethanol solution, remained unchanged under 20% ethanol, and 43.68% of the enzyme activity could still be retained under 30% ethanol. The enzyme has an obvious activation effect at 0-1.5 mol/L NaCl and can tolerate 0.8 mol/L glucose. In conclusion, CdBglA is an acidic and mesophilic enzyme with broad pH stability and strong tolerance to most metal ions, organic solvents, NaCl and glucose. These characteristics may facilitate future theoretical research and industrial production.
beta-Glucosidase ; Sodium Chloride ; Temperature ; Glucose ; Ethanol/chemistry* ; Ions ; Hydrogen-Ion Concentration ; Enzyme Stability ; Substrate Specificity

Chronic refractory wound (CRW) is one of the most challengeable issues in clinic due to complex pathogenesis, long course of disease and poor prognosis. Experts need to conduct systematic summary for the diagnosis and treatment of CRW due to complex pathogenesis and poor prognosis, and standard guidelines for the diagnosis and treatment of CRW should be created. The Guideline forthe diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients ( version 2023) was created by the expert group organized by the Chinese Association of Orthopedic Surgeons, Chinese Orthopedic Association, Chinese Society of Traumatology, and Trauma Orthopedics and Multiple Traumatology Group of Emergency Resuscitation Committee of Chinese Medical Doctor Association after the clinical problems were chosen based on demand-driven principles and principles of evidence-based medicine. The guideline systematically elaborated CRW from aspects of the epidemiology, diagnosis, treatment, postoperative management, complication prevention and comorbidity management, and rehabilitation and health education, and 9 recommendations were finally proposed to provide a reliable clinical reference for the diagnosis and treatment of CRW.