Objective To investigate the mechanism of the interaction between miR-9-5p and tissue metalloproteinase inhibitor 2(TIMP2)on autophagy and apoptosis in multiple myeloma(MM)cells.Methods Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect expression levels of miR-9-5p and TIMP2 in bone marrow samples of 9 patients with newly diagnosed MM and 9 patients with recurrent MM.The correlation of expression levels between the two were analyzed.U266 cells were divided into the miR-control group,the miR-9-5p group,the pcDNA3.1 group,the pcDNA3.1-TIMP2 group,the miR-9-5p+pcDNA3.1 group,and the miR-9-5p+pcDNA3.1-TIMP2 group.The effects of overexpressed miR-9-5p and TIMP2 on autophagy and apoptosis in U266 cells were detected by flow cytometry,immunofluorescence staining and Western blot experiments.The dual luciferase report experiment verified the interaction between miR-9-5p and TIMP2.Results Compared with newly diagnosed MM patients,the expression level of miR-9-5p was increased and the expression level of TIMP2 was decreased in patients with recurrent MM.The expression levels of miR-9-5p and TIMP2 were negatively correlated(P<0.05).Compared with the miR-control group,the miR-9-5p group showed a decrease in the expression level of MAP1LC3B-Ⅱ,an increase in expression levels of MAP1LC3B-Ⅰ and SQSTM1,and a decrease in cell apoptosis rate(P<0.05).Compared with the pcDNA3.1 group,the expression level of MAP1LC3B-Ⅱ was increased in the pcDNA3.1-TIMP2 group,while the expression levels of MAP1LC3B-Ⅰ and SQSTM1 were decreased,and the apoptosis rate of cells increased(P<0.05).Bioinformatics and dual luciferase reporter experiments confirmed that TIMP2 was the target gene of miR-9-5p.Conclusion miR-9-5p inhibits autophagy and apoptosis in MM cells by targeting TIMP2,thereby promoting the occurrence and development of MM.

Objective:To explore the risk factors for death within 1 year after hip fracture surgery in the elderly.Methods:A case control study was made on the clinical data of 551 elderly patients with hip fracture treated in Second Affiliated Hospital of Soochow University from January 2019 to December 2020, including 182 males and 369 females; aged 65-100 years [80(73,86)years]. Joint replacement, cannulated screw fixation or proximal femoral nail fixation were performed. The patients were divided into survival group ( n=494) and death group ( n=57) based on the death within 1 year after surgery recorded at postoperative telephone follow-up. The gender, age, hypertension, diabetes, cardiovascular diseases, chronic respiratory diseases, neurological diseases, chronic renal failure, anemia on admission, fracture types, American anesthesiologist Association (ASA) classification, operative methods, preoperative waiting time, duration of operation and perioperative blood transfusion were recorded in two groups. Univariate Cox regression was used to analyze the correlation between the above indexes and death within 1 year after surgery. All indexes with P<0.2 in the univariate analysis were included in multivariate Cox regression analysis to clarify the independent risk factors for death within 1 year after surgery. Results:Univariate Cox regression analysis showed that death within 1 year after surgery correlated with gender, age chronic respiratory diseases, chronic renal failure and anemia on admission (all P<0.01), but not with hypertension, diabetes, cardiovascular diseases, neurological diseases, fracture types, ASA classification, operative methods, preoperative waiting time, duration of operation or perioperative blood transfusion (all P>0.05). Multivariate Cox regression analysis showed that male ( HR=2.08, 95% CI 1.20, 3.61, P<0.01), age ≥ 80 years ( HR=2.22, 95% CI 1.15, 4.28, P<0.05), chronic respiratory diseases ( HR=2.54, 95% CI 1.19, 5.40, P<0.05), chronic renal failure ( HR=4.57, 95% CI 1.27, 16.44, P<0.05), anemia on admission ( HR=2.82, 95% CI 1.38, 5.76, P<0.01) were significantly associated with death within 1 year after surgery. Conclusion:Male age≥ 80 years, chronic respiratory disease, chronic renal failure and anemia on admission are independent risk factors for death within 1 year after hip fracture surgery in the elderly.

Humans ; Hallux Valgus/surgery* ; Orthopedic Procedures ; Bunion/surgery* ; Joint Diseases/surgery* ; Treatment Outcome

Objective:To investigate the effect of early ambulation time on lower limbs muscle strength and fear of falling in patients with total knee arthroplasty, so as to provide basis for promoting the postoperative rehabilitation of patients undergoing total knee arthroplasty.Methods:A total of 210 patients after unilateral total knee replacement from July 2018 to December 2019 in the Second Affiliated Hospital of Soochow University were included and assigned to experimental group 1, experimental group 2 and experimental group 3 by random digits table method, each group contained 70 cases, and the patients began ambulation at 16, 20, 24 hours after knee replacement, respectively. The peak torque (PT) of knee joints flexors and extensor as well as hamstrings quadriceps ratio (H/Q) were compared among three groups at 1 week, 1 month and 3 months after knee replacement. The fall efficiency was evaluted by Modified Fall Efficacy Scale (MFES), the fear of falling rate was conducted by single item method.Results:After 1 month of knee replacement, the PT of knee joints flexor and extensor and H/Q were (18.73±5.49) N·m, (37.56±7.76) N·m, (48.08±9.19)% and (18.44±5.27) N·m, (37.04±7.07) N·m, (47.49±9.30)% in the experimental group 1 and experimental group 2, which were higher than those in the experimental group 3(16.38±2.85) N·m, (33.75±6.75) N·m, (43.48±7.17)%, the differences were statically significant ( t values were 2.316-3.057, P<0.05). After 1 week of knee replacement, the fear of falling rate were 72.3%(47/65) and MFES scores were (3.14±0.58) points in the experimental group 1, 53.7%(36/67), (3.81±0.65) points and 50.8%(32/63), (3.87±0.74) points in the experimental group 2 and experimental group 3, the fear of falling rate significantly increased and MFES scores significantly decreased in the experimental group 1 compared to the experimental group 2 and experimental group 3, the differences were statistically significant ( χ2 values were 5.780, 4.878, t values were 6.221, 6.129, P<0.05). Conclusions:Twenty hours after knee replacement is the best time for ambulation, which can shortern the time for lower limbs muscle strength recovery and reduce the risk of fear of falling.

Objectives:To investigate the nursing effect of perioperative enhanced recovery after surgery（ERAS）management model in elderly patients with hip fracture.Methods:A retrospective case-control study was conducted to analyze the clinical data of 94 elderly patients with hip fracture admitted to Second Affiliated Hospital of Soochow University from July 2017 to June 2018，including 24 males and 70 females aged 76-98 years［（83.7±5.4）years］. There were 54 patients with intertrochanteric fracture and 40 patients with femoral neck fracture. Surgical method was proximal femoral nail antirotation（PFNA）fixation or artificial hip arthroplasty. A total of 44 patients were managed by ERAS（enhanced recovery group）and 50 patients by traditional nursing（traditional rehabilitation group）. The off-bed time was compared between the two groups from aspects of different fracture sites，bone mineral density and causes of injury. The visual analogue scale（VAS）was tested preoperatively and at postoperative days 1，3 and 7. The complications were recorded at postoperative 1 month. The activity of daily living（ADL）score was assessed preoperatively and at postoperative 1 week，1 month and 3 months. The Harris hip score was assessed at postoperative 1 week，1 month，3 months and at the last follow-up. The length of hospital stay and death at postoperative 3 months and at the last follow-up were recorded.Results:All patients were followed up for 12-24 months［（17.7±6.2）months］. In enhanced recovery group，the off-bed time of patients with different fracture sites（femoral neck，femoral intertrochanter），bone mineral density（>-2.5 SD，≤-2.5 SD）and causes of injury（falls，traffic accidents and others）were markedly shortened as compared with traditional rehabilitation group（ P<0.01）. There was no significant difference in VAS between the two groups before operation（ P>0.05）. However，the VAS in enhanced recovery group was（3.4±0.9）points，（2.7±0.5）points，（1.7±0.6）points at postoperative days 1，3 and 7，significantly lower than that in traditional rehabilitation group［（4.3±1.1）points，（3.5±0.5）points，（2.7±0.9）points］（ P<0.01）. One month after operation，the incidence of pulmonary infection was 0% in enhanced recovery group and 28%（14/50）in traditional rehabilitation group（ P<0.01）. While there showed no significant differences in incidences of cardiovascular system，nervous system，urinary tract infection，lower limb deep vein thrombosis，impaired liver and kidney function，anemia，electrolyte disorder or hypoalbuminemia between the two groups（ P>0.05）. There was no significant difference in preoperative ADL score between the two groups（ P>0.05）. The ADL score in enhanced recovery group was（37.0±6.6）points，（70.1±8.4）points，（86.2±9.3）points at postoperative 1 week，1 month and 3 months，significantly higher than that in traditional rehabilitation group［（26.5±10.4）points，（50.1±11.4）points，（70.7±9.0）points］（ P<0.01）. The Harris hip score in enhanced recovery group was（80.9±8.6）points at postoperative 1 month，significantly higher than that in traditional rehabilitation group［（71.1±9.2）points］（ P<0.01）. There were no significant differences in Harris hip score between the two groups at postoperative 1 week，3 months and at the last follow-up（ P>0.05）. The length of hospital stay was（4.7±2.4）days in enhanced recovery group，significantly shorter than（7.8±3.9）days in traditional rehabilitation group（ P<0.01）. The enhanced recovery group showed no death within 3 months after operation and 2 deaths［5%（2/44）］at the last follow-up，while the traditional rehabilitation group revealed 4 deaths［8%（4/50）］within 3 months after operation. There was no significant difference in the mortality between the two groups until the last follow-up（ P>0.05）. Conclusion:For elderly patients with hip fracture，perioperative ERAS management model can effectively shorten periods in bed，relieve postoperative pain，reduce incidence of postoperative pulmonary infection，accelerate recovery of hip function，improve quality of life，shorten length of hospitalization and promote early recovery.

Objective:To investigate the role of macrophages (M?) in the pathogenesis of modified immune-mediated aplastic anemia (AA) mice model.Methods:Before the establishment of the F1 AA mice model by total-body irradiation combined with allogeneic lymphocyte infusion, the mice of the CLO+AA group were treated with clodronate (CLO) liposomes to remove macrophages, and those of the PBS+AA group were treated with phosphate-buffered saline (PBS) liposomes and used as control. The severity of AA was observed by bone marrow (BM) pathological examination and peripheral blood cell count. Flow cytometry (FCM) was used to detect the CD4 +/CD8 + T lymphocyte subsets in the BM and M? subsets in the BM and spleen of each group. The levels of IFN-γ, TNF-α, G-CSF, GM-CSF, EPO, and TPO in the peripheral blood were detected using enzyme-linked immunosorbent assay. Finally, the relationships between inflammatory factors and M? subsets were analyzed. Results:The BM fatty conversion of mice in the CLO+AA group was significantly alleviated compared with the PBS+AA group. Hemoglobin counts were (91.50±31.63) and (110.65±24.15) g/L, respectively, and the platelet counts were (90.85±121.90) × 10 6/L and (461.13±483.45) ×10 6/L, respectively. The differences were all statistically significant (all P<0.05) . After removing macrophages, the proportions of CD4 + and CD8 + T lymphocytes in BM of mice in the CLO+AA group decreased, but the reduction of CD8 + T cells was more significant. The proportions of CD4 + T cells and CD8 + T cells in BM of the PBS+AA group were (18.5±10.17) % and (36.23±6.40) %, respectively, and in the CLO+AA group were (7.58±8.00) % and (6.67±5.78) %, respectively. Similarly, the percentage of macrophages in the spleen and BM in the CLO+AA group was significantly reduced compared with the PBS+AA group, most of which were M1 macrophages ( P<0.05) . The levels of IFN-γ in peripheral blood of the PBS+AA and CLO+AA groups were (602.37±104.62) ng/L and (303.01±87.22) ng/L, respectively, the levels of TNF-α were (34.46±1.42) ng/L and (23.25±4.21) ng/L, respectively, the levels of GM-CSF were (9.32 ± 2.00) ng/L and (64.85±12.25) ng/L, respectively, the levels of G-CSF were (5 891.78±2 632.39) ng/L and (17 784.16±488.36) ng/L, respectively, the levels of EPO were (9 667.31±4 501.95) ng/L and (2 078.02±897.56) ng/L, respectively, and the levels of TPO were (6.36±2.09) ng/L and (11.67±2.86) ng/L, respectively (all P<0.05) . Conclusions:This study confirmed that macrophages were involved in the pathogenesis of AA, and the degree of BM damage in AA mice was improved by removing macrophages in advance. The imbalance of M1/M2 macrophages and the changes of IFN-γ and TNF-α may be important mechanisms that eventually lead to AA.

Bone Cements ; Calcium ; Calcium Phosphates ; Materials Testing

Objective To observe the effect of a hypoxia mimicking agent deferoxamine (DFO) on the mineral density,volume,architecture,strength,and metabolism of the bones in type 1 diabetic mice withosteoporosis.Methods Type 1 diabetic mice model was established by intraperitoneal injections of streptozotocin.The mice were divided into control (normal mice),diabetes mellitus,and DFO groups.Micro-CT was used to analyze the bone mineral density,volume,architecture,and strength of the trabecule in the distal part of femurs.Three point bending test was carried out to evaluate the bone strength.Hematoxylin and eosin (HE) staining was performed to observe the alteration in the number of osteoblasts.Real-time PCR was used to detect the mRNA expressions of Runt-related gene 2 (Runx-2),osteoclacin,and tartrate resistant acid phosphatase (TRAP) in tibias.Western blot was used to detect the protein expressions of Hypoxia-inducible factor-1α(HIF-1α) and vascular endothelial growth factor (VEGF) in tibias.Results There was a decrease in mineral density,volume,strength of bones as well as deteriorated trabecular microarchitecture in diabetic mice as compared to control mice,which were partially improved by DFO treatment.Moreover,DFO treatment increased the number of osteoblasts and mRNA expression levels of Runx-2,osteoclacin,TRAP,as well as protein expression levels of HIF-1 α and VEGF(P＜0.05).Conclusion Bone loss could be partially prevented by DFO treatment in type 1 diabetic osteoporosis mice,which might be ascribed to increased bone formation via stimulating hypoxia inducible factor singnaling pathway.

Objective: To explore the blood lead level and its relationship with behavior in school-age children from rural areas of Chongqing. Methods: A total of 697 students from grades 3 to 6 in the fall semester of 2014 from 14 rural townships in one district of Chongqing was selected by using the random cluster sampling method. Blood were sampled to analyze the lead level. Neurobehavioral tests were performed to determine their personal cognitive and memory ability. Questionnaires and physical examinations were administered to obtain the information of confounding factors. All students were divided into Q₁-Q₄ groups according to the quartile of their blood lead level. The relationship between the blood lead level and behavior was analyzed by multivariate logistic regression model and restricted spline regression model. Results: The mean age of 697 students was (10.07±1.36) years old, and the median (interquartile range) of their blood lead level was 44.31 (35.42) μg/L. Multivariate logistic regression model showed that after adjusting for age, gender, body mass index and maternal culture level, compared with Q₁ group, the OR (95%CI) values of high digit symbol substitution test (DSST) scores and high overall memory quotient (MQ) scores in Q₃ group were 1.65 (1.01-2.70) and 2.10 (1.21-3.62), and the OR (95%CI) value of high long term memory (LTM) scores in Q₄ group was 0.53 (0.31-0.92). The results of the restricted spline regression model showed that the dose-response curves between the blood lead level and MQ/LTM test scores were both parabolic (P<0.05). Conclusion The blood lead level of school-age children from rural areas of Chongqing is the same as that from other areas of China, but slightly higher than that from other areas of Chongqing. Children with higher blood lead level have poor long-term memory ability.

Objective To explore the differences in autophagic expression levels between hypertrophic scar (HS) tissue and normal skin tissue,and further investigate the relationship between hypertrophic scar formation and autophagy protein expression through the rabbit ear hypertrophic scar model.Methods 30 patients with hypertrophic scar were collected.One hypertrophic scar tissue and one normal skin tissue were harvested.The relative expressions of LC3,P62 and Beclin-1 in each tissue specimen were detected by immunohistochemistry and Western blot.Western blot was used to detect the autophagic-associated protein LC3 (MAPLC3),P62 and Beclin-1 in the hypertrophic scar tissue of rabbit ear and the corresponding normal tissue of rabbit ears at 4 weeks,8 weeks,12 weeks,and 24 weeks,and further explore their clinical significance.Results In vivo,the expression of hypertrophic scar tissue protein LC3 and Beclin-1 was significantly stronger than that in normal skin tissue (P ＜ 0.05).The expression of P62 was significantly weaker than that in normal skin tissue (P ＜ 0.05).In animal experiments,during the process of HS formation,the protein expression of LC3 gradually increased,while the protein expression of P62 gradually decreased;the protein expression of Beclin-1 was higher than that of normal rabbit ears tissue,with statistically significant differences (P ＜ 0.05).Conclusions The expression of LC3 and Beclin-1 in human hypertrophic scar tissues is higher than that in normal tissues.While the expression of P62 is lower than that in normal tissues.That is,the expression of autophagy in human hypertrophic scar tissue showed an upward trend in a certain period of time,and was significantly higher than that in normal tissue.