Objective To evaluate the effect of tissue-engineered nerve graft combined with tetramethylpyrazine in the repair of ischiadic nerve defect in rats.Methods Fifty-five adult SD rats were allocated into groups A (n =15),B (n =15),C (n =15) and D (n =10),according to the random number table.All experimental sciatic nerves were at the right hind side.After the rats were anesthetized with 10 g/L pentobarbital through abdominal injection,a 1.5 cm nerve sciatic nerve was excised at the point 0.5 cm away from the lower margin of the piriformis.Four grafts were used to bridge the nerve defect,including tissue-engineered nerve transplanted neural stem cells (NSCs) that were cultured in medium containing tetramethylpyrazine for 1 week (group A),tissue-engineered nerve transplanted NSCs that were cultured in tetramethylpyrazine-free medium for 1 week (group B),acellular nerve allograft (group C),and nerve autograft (group D).After operation,group A was administered tetramethylpyrazine for 12 weeks in the operative site.For other groups,the same volume of normal saline was used instead of tetramethylpyrazine.Repair results were detected with the sciatic function index (SFI),nerve electrophysiological evaluation,fluorescent microscopy,horseradish peroxidas (HRP) retrograde tracer and nerve fiber regeneration study.Results At postoperative 2 weeks,no differences were found in SFI and electrophysiological evaluation among the groups,there was an intensive fluorescence in groups A and B and no fluorescence in group C,and large nestin-positive cells were observed in groups A and B.At postoperative 12 weeks,SFI was-17.52 ± 2.41 in group A,-25.74 ± 2.85 in group B,-36.12 ± 3.41 in group C and-15.87 ± 2.26 in group D;electrophysiological evaluation showed nerve conduction velocity of (9.43 ± 0.40) m/s in group A,(7.76 ± 0.31) m/s in group B,(5.87 ± 0.67) m/s in group C and (10.16 ± 0.39) m/s in group D;fluorescence microscope and histological staining showed NSCs could survive in vivo,migrate and differentiate into neurons and gliocytes;HRP-positive neurons were 885.40 ± 19.91 in group A,684.57 ± 38.37 in group B,390.33 ± 43.41 in group C and 941.67 ± 32.54 in group D;regenerated nerve fibers,often myelinated were large in group A.All the measures in group A were better compared to groups B and C (P ＜ 0.05),but didn't differ from those in group D (P ＞0.05).Conclusion Tissue-engineered nerve transplanted NSCs cultured in medium including tetramethylpyrazine can effectively repair rat sciatic nerve defect,and promote peripheral nerve regeneration and lower limb motor function recovery.

BACKGROUND:Acellular nerve scaffolds have the three-dimensional structure of natural nerves and low immunogenicity,but their effect on long nerve defects is still not ideal.Therefore,it is necessary to construct tissueengineered nerve using acellular nerve and seed cells in order to improve the therapeutic effect.OBJECTIVE:To systemically review the efficacy of combination of acellular nerve grafts (ANGs) and mesenchymal stem cells (MSCs) or Schwann cells (SCs) transplantation in the treatment of sciatic nerve defects in a rat model.METHODS:Randomized controlled trials (RCTs) about the effects of combination of ANGs and MSCs or SCs transplantation for sciatic nerve defects in rats were searched in PubMed,The Cochrane Library,EMbase,CNKI,WanFang and VIP from inception to July 2016.Three reviewers independently screened literature according to the inclusion and exclusion criteria,extracted data,and assessed the risk of bias of included studies.Then,a Meta-analysis was performed using Review Manger5.3 software.RESULTS AND CONCLUSION:A total of 10 RCTs involving 252 rats were included.The results of meta-analysis showed that:compared with the control group (simple acellular nerve scaffold group),the sciatic functional index (SFI) of the combined group (combination of ANGs and MSCs or SCs transplantation) were superior at 2 weeks [SMD=2.73,95％ CI (1.92,3.45),P ＜ 0.000 01],4 weeks [SMD=4.57,95％ CI (3.43,5.70),P ＜ 0.000 01],6 weeks [SMD=1.62,95％CI (0.18,3.06),P=-0.03],8 weeks [SMD=4.90,95％ CI (2.96,6.84),P ＜ 0.000 01] after surgery.The nerve conduction velocity [SMD=1.39,95％ CI (0.99,1.78),P ＜ 0.000 01),latency period (MD=-0.98,95％ CI (-1.19,-0.76),P ＜ 0.000 01],and amplitude [SMD=1.23,95％ CI (0.62,1.85),P ＜ 0.000 1] were superior at 12 weeks after surgery.The myelin sheath thickness was superior at 8 weeks [MD=0.14,95％ CI (0.07,0.21),P ＜ 0.000 1],12 weeks [SMD=1.85,95％ CI (1.63,2.08),P ＜ 0.000 01] and the number of myelinated nerve fibers were superior at 12 weeks [SMD=3.59,95％CI (2.63,4.55),P ＜ 0.000 01] after surgery.The gastrocnemius wet weight was superior at 8 weeks after surgery [SMD=4.22,95％ CI (2.40,6.03),P ＜ 0.000 01].Current evidence indicates that the combination of ANGs and MSCs or SCs can promote the regeneration and functional recovery of the peripheral nerve.Due to the limited quality of the included studies,the above conclusion should be verified by conducting high-quality and large-scale RCTs.

BACKGROUND: Ipsilateral femoral shaft and neck fracture is a rare clinical injury, and its treatment has attracted more and more attention from scholars. At present, there are many internal fixation methods. OBJECTIVE: To retrospectively analyze efficacy of three internal fixation methods in the treatment of ipsilateral femoral shaft and neck fracture, and its indications. METHODS: Thirty-three cases of ipsilateral femoral neck and shaft fracture treated by three different internal fixation implants between January 2015 and September 2018 were retrospectively studied. Patients were divided into three groups according to different internal fixation: intramedullary nail only group (group A; n=11), retrograde nail with cannulated screw group (group B; n=10) and plate with cannulated screw group (group C; n=12). The general clinical data, the intraoperative situations, the postoperative follow-up conditions and complications were compared among the three groups. RESULTS AND CONCLUSION: (1) There was no significant difference between the three groups with regard to age, gender, cause of injury, AO fracture classification, Garden classification and average time from injury to surgery (P > 0.05). (2) There were statistically significant differences among the three groups in incision length (P < 0.05); no significant difference in incision length was found between the groups A and B (P > 0.05); the incision length was smaller in groups A and B than in group C (P < 0.05). There were no significant differences among the three groups in operation time and intraoperative blood loss (P > 0.05). (3) There were no significant differences among the three groups in time to bony union in both femoral shaft and neck, complication rate, and excellent and good rate of Harris hip score (P > 0.05). (4) Femoral neck fracture delayed healing occurred in one case of the group A; re-fracture occurred in one case of the group B; and the fracture end was located under the femoral tuberosity; screw loosening occurred in one case of the group C. (5) It is indicated that all three internal fixation methods can achieve good therapeutic effects. The operator can choose the skilled internal fixation method according to the specific conditions of the patient.

Objective: To explore the relationship between adolescents physical fitness and depressive symptoms, and to provide reference for the early prevention and intervention of depressive symptoms and improvement of physical fitness in Chinese adolescents. Methods: From September to December 2021, a total of 8 102 adolescents were selected by random cluster sampling method in Shanghai, Urumqi, Changsha and Kunming. The Center for Epidemiologic Studies Depression Scale(CES-D) was used to investigate the depressive symptoms, and completed grip strength, standing long jump, 50 m running, modified sitting forward flexion, 20 s repeated traverse, 30 s sit ups, 20 m round trip running (20 m SRT) test. χ 2 test, Goodman Kruskal Gamma and Logistic regression analysis were used to analyze the relationship between physical fitness index(PFI) and depressive symptoms. Results: The overall detection rate of depressive symptoms in adolescents with high level PFI was 23.4%, and the detection rate of low level adolescents was 26.3%, with a statistically significant difference ( χ 2=6.73, P =0.01). There was a significant positive correlation between PFI and depressive symptoms in the high school group ( G=0.09, P <0.05) and the boy group ( G=0.12, P < 0.05 ), and there was no significant association between PFI and depressive symptoms in the junior high school group and the girl group ( P >0.05). After adjusting for gender and age in the Logistic regression model, compared with those with high PFI, the risk of depressive symptoms in those with low PFI was 1.18 times (95% CI =1.05-1.33). Conclusion There is a correlation between physical fitness and depressive symptoms in adolescents. Adolescents with low PFI are at higher risk of developing depressive symptoms than those with high PFI.

Objective To explore the molecular mechanism by which FEZF1-AS1 overexpression promotes progression of non-small cell lung cancer(NSCLC)via the miR-130a-5p/CCND1 axis.Methods TCGA database was used to analyze FEZF1-AS1 expression levels in NSCLC.FEZF1-AS1 expression was detected by qRT-PCR in clinical specimens of NSCLC tissues and NSCLC cell lines,and its correlation with clinical features of the patients were analyzed.The binding sites of FEZF1-AS1 with hsa-miR-130a-5p and those of hsa-miR-130a-5p with CCND1 were predicted.CCK8 assay,clone formation assay,scratch assay,and Transwell assay were employed to examine the effects of FEZF1-AS1 knockdown and hsa-miR-130a-5p inhibitor on proliferation,invasion,and migration abilities of lung cancer cell lines.Dual luciferase assay was used to verify the binding of FEZF1-AS1 with hsa-miR-130a-5p and the binding of hsa-miR-130a-5p with CCND1.Western blotting was performed to detect the changes in CCND1 protein expression level in H1299 and H358 cells following FEZF1-AS1 knockdown and treatment with hsa-miR-130a-5p inhibitor.Results FEZF1-AS1 was highly expressed in NSCLC tissues in close correlation with lymph node metastasis and also in H1299 and H358 cell lines(all P<0.05).FEZF1-AS1 knockdown obviously reduced proliferation,migration,and invasion abilities of NSCLC cells(P<0.05).Dual luciferase assay confirmed the binding of hsa-miR-130a-5p with FEZF1-AS1 and CCND1(P<0.05),and hsa-miR-130a-5p inhibitor significantly inhibited proliferation,migration,and invasion of NSCLC cells(P<0.05).FEZF1-AS1 knockdown significantly reduced CCND1 protein expression in NSCLC cells,and this effect was strongly inhibited by treatment with hsa-miR-130a-5p inhibitor(P<0.05).Conclusion FEZF1-AS1 is highly expressed in NSCLC tissue in close correlation with lymph node metastasis to promote cancer progression through the miR-130a-5p/CCND1 axis.

BACKGROUND:Pauwels Ⅲ femoral neck fracture is a typical unstable fracture characterized by high vertical shear force and high incidence of postoperative complications.At present,there are many fixation methods for Pauwels Ⅲ fracture,and there is no clear conclusion as to which internal fixation is the best fixation method for Pauwels Ⅲ femoral neck fracture in young adults. OBJECTIVE:To compare the clinical effect of three inverted triangle cannulated screws assisted transverse lag screws and femoral neck system in fixing Pauwels Ⅲ femoral neck fractures of young adults. METHODS:From May 2021 to December 2022,21 young and middle-aged patients with Pauwels Ⅲ femoral neck fracture were treated with three inverted triangle cannulated screws assisted by transverse lag screws and femoral neck system in Affiliated Hospital of Southwest Medical University.Of them,9 patients were treated with three inverted triangle cannulated screws and one transverse lag screws perpendicular to the fracture line as the 3+1 cannulated screw group and 12 patients were treated with femoral neck system as the femoral neck system group.The two fixation methods were compared in terms of operation time,intraoperative blood loss,total incision length,intraoperative fluoroscopy times,fracture healing time,and limb function. RESULTS AND CONCLUSION:(1)All patients were followed up.Patients in the 3+1 cannulated screw group were followed up for 10-25 months,with a mean of(17.44±4.30)months.The patients in the femoral neck system group were followed up for 8-24 months,with a mean of(15.58±4.68)months.(2)The intraoperative fluoroscopy times and Harris score at 3 months postoperatively in the femoral neck system group were better than those in the 3+1 cannulated screw group,and the difference was statistically significant(P<0.05).The intraoperative blood loss,total incision length,and femoral neck shortening distance in the 3+1 cannulated screw group were better than those in the femoral neck system group,and the differences were statistically significant(P<0.05).There was no significant difference in operation time,fracture healing time,and Harris score at the last follow-up between the two groups(P>0.05).(3)It is indicated that three inverted triangle cannulated screws assisted transverse lag screw and femoral neck system can achieve good clinical effects in the treatment of young and middle-aged Pauwels Ⅲ femoral neck fracture.The femoral neck system has fewer intraoperative fluoroscopy times and better early overall stability,while the 3+1 cannulated screw is more minimally invasive,easier to operate.Both fixation methods are worthy of clinical application and promotion,and can be selected according to the actual clinical situation.

Objective To explore the molecular mechanism by which FEZF1-AS1 overexpression promotes progression of non-small cell lung cancer(NSCLC)via the miR-130a-5p/CCND1 axis.Methods TCGA database was used to analyze FEZF1-AS1 expression levels in NSCLC.FEZF1-AS1 expression was detected by qRT-PCR in clinical specimens of NSCLC tissues and NSCLC cell lines,and its correlation with clinical features of the patients were analyzed.The binding sites of FEZF1-AS1 with hsa-miR-130a-5p and those of hsa-miR-130a-5p with CCND1 were predicted.CCK8 assay,clone formation assay,scratch assay,and Transwell assay were employed to examine the effects of FEZF1-AS1 knockdown and hsa-miR-130a-5p inhibitor on proliferation,invasion,and migration abilities of lung cancer cell lines.Dual luciferase assay was used to verify the binding of FEZF1-AS1 with hsa-miR-130a-5p and the binding of hsa-miR-130a-5p with CCND1.Western blotting was performed to detect the changes in CCND1 protein expression level in H1299 and H358 cells following FEZF1-AS1 knockdown and treatment with hsa-miR-130a-5p inhibitor.Results FEZF1-AS1 was highly expressed in NSCLC tissues in close correlation with lymph node metastasis and also in H1299 and H358 cell lines(all P<0.05).FEZF1-AS1 knockdown obviously reduced proliferation,migration,and invasion abilities of NSCLC cells(P<0.05).Dual luciferase assay confirmed the binding of hsa-miR-130a-5p with FEZF1-AS1 and CCND1(P<0.05),and hsa-miR-130a-5p inhibitor significantly inhibited proliferation,migration,and invasion of NSCLC cells(P<0.05).FEZF1-AS1 knockdown significantly reduced CCND1 protein expression in NSCLC cells,and this effect was strongly inhibited by treatment with hsa-miR-130a-5p inhibitor(P<0.05).Conclusion FEZF1-AS1 is highly expressed in NSCLC tissue in close correlation with lymph node metastasis to promote cancer progression through the miR-130a-5p/CCND1 axis.

BACKGROUND:It has been found that abnormal apoptosis of bone tissue cells induced by abnormal iron metabolism plays an important role in the progression of osteoporosis. OBJECTIVE:To investigate the effect of naringin on iron metabolism and cell apoptosis in bone tissue of rats with osteoporosis. METHODS:Fifty 2-month-old female Sprague-Dawley rats were randomly divided into five groups with 10 rats in each group:sham group,osteoporosis group,naringin low-dose group,naringin high-dose group,and naringin high-dose+DKK-1 group.Except for the sham group,rat models of osteoporosis were established by removing bilateral ovarian tissues in the other groups.At 8 weeks after modeling,rats in the naringin low-and high-dose groups were given 100 and 400 mg/kg/d naringenin by gavage,respectively,and rats in the naringenin high dose+DKK-1 group were given 400 mg/kg/d naringin by gavage and subcutaneous injection of 25 mg/kg/d DKK-1,an inhibitor of the Wnt1 signaling pathway,for 7 consecutive days.Relevant indexes were detected after administration. RESULTS AND CONCLUSION:Compared with the osteoporosis group,naringin could enhance the bone mineral density and serum calcium and superoxide dismutase levels in rats(P<0.05),and reduce the serum levels of osteocalcin,malondialdehyde,and phosphorus(P<0.05),while DKK-1 could partially inhibit the interventional effect of naringin(P<0.05).Results from Micro-CT scanning,hematoxylin-eosin and TUNEL staining showed that compared with the osteoporosis group,naringin significantly improved bone microstructure and reduced the rate of cell apoptosis,while DKK-1 partially inhibited the interventional effect of naringin.Immunofluorescence staining results showed that compared with the osteoporosis group,naringin could reduce the oxygen content,anti-tartaric acid phosphatase expression,and elevate the expression of alkaline phosphatase in active tibia tissues(P<0.05),while DKK-1 could partially inhibit the interventional effect of naringin(P<0.05).Results from Prussian blue staining and immunohistochemical staining showed that compared with the osteoporosis group,naringin reduced iron deposition in bone and liver tissues as well as the expression of transferrin receptor 1(P<0.05),and elevated the protein expression of ferroportin 1(P<0.05)in bone tissue,and DKK-1 partially inhibited the intervention of naringin(P<0.05).PCR and western blot assay of tibia specimens showed that compared with the osteoporosis group,naringin decreased the expression of anti-tartrate acid phosphatase,transferrin receptor 1 and Bax(P<0.05),and elevated the expression of alkaline phosphatase,ferroportin 1,Bcl-2,Wnt1 and β-catenin(P<0.05),while DKK-1 partially inhibited the interfering effect of naringin(P<0.05).To conclude,naringin inhibits the progression of osteoporosis by reducing iron deposition and apoptosis rate in bone tissue,which may be related to the activation of the Wnt1 signaling pathway.

Objective To observe biomechanical characteristics and advantages of the self-developed anatomical locking plate of sternoclavicular joint by comparison with the radial distal oblique T-shaped locking plate and sternoclavicular hook plate. Methods Nine embalming and moistening adult corpses were selected, including 6 males and 3 females. Model of complete dislocation of the sternoclavicular joint caused by complete removal of the sternoclavicular joint specimen. The bilateral sternoclavicular joints of 9 specimen models were randomly numbered, matched and divided into Group A(experimental group, sternoclavicular joint anatomical locking plate), Group B(control group 1, the radial distal oblique T-shaped locking plate) and Group C(control group 2, sternoclavicular hook plate), with 6 sternoclavicular joints in each group. All specimens were placed with steel plates on both sides and fixed on the universal mechanical test machine. Three biomechanical experiments were carried out, including loading of distal clavicle, torsion of distal clavicle and anti-pull out of sternum handle screw. Results Distal clavicle loading test: the load-displacement of specimens in three groups showed a linear relationship. The compressive deformation resistance in experimental group was stronger than that in two control groups. Distal clavicle torsion test: the relationship between torque and torsion angle was linear. The torsional deformation resistance in experimental group was stronger than that in two control groups. Anti-pullout test of sternum handle screw: there was a significant difference in the maximum anti-pullout force of sternum handle screw among the three groups (P＜0.05). The anti-pullout performance of the sternum handle screw in the experimental group was better than that in the two control groups. Conclusions The self-developed sternoclavicular joint anatomical locking plate is superior to the oblique T-shaped locking plate of distal radius and the plate of sternoclavicular hook in terms of anti-compression, anti-torsion and anti-pullout of sternoclavicular screw, so as to provide an ideal internal fixation device for the treatment of fracture and dislocation of sternoclavicular joint.

Biological sample pretreatment is an important step in biological sample analysis. Due to the diversity of biological matrices, the analysis of target substances in these samples presents significant challenges to sample processing. To meet these emerging demands on biopharmaceutical analysis, this paper summarizes several new techniques of on-line biological sample processing: solid phase extraction, solid phase micro-extraction, column switching, limited intake filler, molecularly imprinted solid phase extraction, tubular column, and micro-dialysis. We describe new developments, principles, and characteristics of these techniques, and the application of liquid chromatography-mass spectrometry (LC-MS) in biopharmaceutical analysis with these new techniques in on-line biological sample processing.