Objective To construct lentiviral vectors containing small interfering RNA (siRNA) sequence of Siglec-1 and to screen the effective vector.Methods Three fragments of Siglec-1 siRNA were designed and cloned into pGCSIL-GFP lentiviral plasmid.And then the plasmid was cotransfected into 293T cells with pHelper 1.0 and pHelper 2.0 plasmids.Forty-eight hours later,culture supernatant with virus particles was collected and concentrated.Virus titer was determined by 10-fold serial dilution method and virus was transduced into primary cultured mouse bone marrow-derived macrophages (BMM).Flow cytometry and QRT-PCR were used to screen effective vector with inhibition ability.Results Three vshRNA lentiviral plasmids and a control plasmid were constructed successfully and verified by DNA sequencing.Virus titer was between 1×10s TU/ml and 1×109 TU/ml,which was suitable for in vitro and in vivo experiments.The Lv-1 could inhibit Siglec-1 expression effectively in vitro transduction of BMM.Conclusion Lentiviral vectors containing siRNA sequence of Siglec-1 were constructed successfully and an effective vector was screened,which may lay the foundation for using the vector in gene knockdown experiment in vivo.

Objective By in vitro culture of mouse macrophage cell line RAW264. 7 and primary mouse bone marrow macrophages, the expression of Siglec-1 when stimulated by ox-LDL was observed. Meanwhile, Siglec-1 was up-regulated by M-CSF and down-regulated by small interference RNA targeting Siglec-1 ( si-RNA-Siglec-1) , and the expression of chemokines and lipid uptake ability by macrophages were observed, to explore the role of Siglec-1 on macrophages in atherosclerosis. Methods LDL was oxidized by copper. According to preliminary experiment results, ox-LDL 100 μg/ml was selected as a stimulus. There were 6 experimental groups:normal control group,ox-LDL 100 μg/ml group, ox-LDL 100 μg/ml + si-RNA 2509 2 ng/ml group,ox-LDL 100 μg/ml + si-RNA 3618 2 ng/ml group,ox-LDL 100 μg/ml + M-CSF 5 ng/ml group and ox-LDL 100 μg/ml + M-CSF 10 ng/ml group. si-RNA-Siglec-1 was transfected into macrophage to inhibit the expression of Siglec-1, whereas M-CSF 10 ng/ml or 5 ng/ml were added into the culture medium to enhance the expression of Siglec-1. Quantitative real-time polymerase chain reaction ( qRT-PCR) was used to determine the interfere efficiency of si-RNA-Siglec-1 or M-CSF. After stimulation with ox-LDL for 48 h, cell culture supernatants were collected to determine MIP-1 alpha, MCP-1 and IL-8 concentration by ELISA (n =3 for each group) to evaluate the activation of macrophages. Internalization of lipid particles by macrophages was analyzed by oil red 0 staining. Results Observed by fluorescence microscope, si-RNA-Siglec-1 could be effectively transfected into macrophages with a transfection efficiency about 90% ;PCR results showed that si-RNA 2509 and si-RNA 3618 in a concentration of 40 pmol/L had an inhibition rate of 0. 54 ±0. 11 or 0. 52 ±0. 16 vs 1. 00 ±0. 24 (control group) , t =5. 227 and 4. 992, respectively, all P < 0.01, while M-CSF 10 ng/ml could increase Siglec-1 mRNA expression approximately 4-fold (4. 16 ± 1. 25 vs 1.00 ±0. 24, t =7. 448, P<0. 01). The secretion of MCP-1, MIP-1 alpha, and MIP-2 in si-RNA3618-Siglec-1 group [(359. 28±47. 80) pg/ml, (33. 76 ± 14. 28) ng/ml and (7.87±1.55) ng/ml for MCP-1,MIP-1 alpha, and MIP-2, respectively] was significantly reduced in compare with ox-LDL 100 μg/ml group [ (577. 89 ± 35. 95 ) pg/ml, (69. 17 ± 11. 82) ng/ml and (12.28 ± 1.19) ng/ml for MCP-1, MIP-1 alpha, and MIP-2, respectively], with P value of 0.01, 0.05 and 0.01. In contrary, ox-LDL 100 μg/ml plus M-CSF 10 ng/ml group could significantly promote macrophage chemokine secretion [ (672. 89 ± 43.80) pg/ml, (101.31 ±24.17) ng/ml and (14.81 ±0.54) ng/ml for MCP-1, MIP-1 alpha, and MIP-2, respectively], with P < 0.05 compared with ox-LDL 100 μg/ml group. Meanwhile, lipid intemalization and foam cell formation was inhibited in si-RNA3618-Siglec-l group while ox-LDL 100 μg/ml plus M-CSF 10 ng/ml group could enhance the phagocytosis of ox-LDL by macrophage. Conclusions Siglec-1 may served as a potential phagocytic receptor for ox-LDL involving in macrophage uptake of lipid and turn into foam cells. Furthermore, it can active macrophages and enhance the secretion of MIP-1 alpha, MCP-1 and IL-8, attracting more macrophages and lymphocytes to the site of inflammatory plaque. Targeted inhibition of Siglec-1 reduces macrophage uptake of lipid and secretion of chemokines. Siglec-1 may possibly serve as a potential target of treatment or delay the development of atherosclerosis.

Severe rotating maxillary central incisors were aligned by NiTi segmental arch,then 2 ×4 technique was applied to correct rota-ted teeth.The treatment was effective and convenient.

BACKGROUND:Indian hedgehog homolog (Ihh) protein and its signal protein Gli1, as wel as Runt related transcription factor 2 (Runx2) are closely related to the pathogenesis of osteoarthritis. The increased expression of these factors is one of the major causes of degenerative joint changes.
OBJECTIVE:To explore the roles of Ihh, Gli1 and Runx2 in the development of osteoarthritis.
METHODS:Thirty Sprague-Dawley rats were randomly divided into two groups:control group (n=10) and model group (n=20). In the model group, rats received unilateral anterior cruciate ligament transection, to establish osteoarthritis model. Ten experimental rats were kil ed at 4 and 12 weeks after surgery respectively. Another 10 rats received unilateral knee arthrotomy as pseudo-operation controls and 10 pseudo-operation rats were kil ed at 12 weeks after surgery.
RESULTS AND CONCLUSION:In the model group, cartilage degeneration was obvious at 4 weeks and became severer at 12 weeks after anterior cruciate ligament transection operation. Expression levels of Ihh, Gli1 and Runx2 in the cartilage were increased significantly at 4 weeks after operation, but decreased at 12 weeks after operation. Experimental findings indicate that Ihh, Gli1 and Runx2 play important roles in the development of osteoarthritis, and their expression levels are significantly increased at early stage of osteoarthritis, which can be regarded as the indicators in the prophylaxis and treatment research of osteoarthritis.

Objective To investigate the influencing factors of the indwelling catheter time in patients with prostate hyperplasia and malignant tumor after prostate surgery.Methods A total of 447 prostate surgery patients with prostate hyperplasia and malignant tumor were selected for medical record review using convenience sampling method. The demographic data,diagnose,complications,surgical method,medical advice and nursing care items were collected. The indwelling catheter time in or outward of the hospital and its influence factors were analyzed.ResultsA number of 300(44.74%) patients had urination disorders before operation.A number of 258(57.72%) patients had complications. Little information about rehabilitated exercise of urination function was recorded.There were statistically significant differences in indwelling catheter time in the hospital among benign prostatic hyperplasia patients with different education levels, health payments and complications (t/F=5.048, -2.433, 2.086;P<0.05). Significant differences in indwelling catheter time outward of the hospital were also observed in patents with different preoperative urination disorders (t=0.551, P<0.05). Prostate malignancy patients with different resident places and bad habits had statistically significant differences in indwelling catheter time outward of the hospital (t=2.062, -1.974;P<0.05). For patients with benign prostatic hyperplasia, education levels, health payment and complications could explain 60.8% of changes in indwelling catheter time in the hospital. For patients with prostate malignancy, resident place and bad habits could explain 68.4% of changes in indwelling catheter time outward of the hospital. According to medical records, only 42.5% patients had urinary flow rate, urodynamic and residual urine volume examinations. Only 17.4% of medical records had urinary function related admission, which were found in 26.4% of medical records when patients were discharged.ConclusionsMedical workers are lack of attentions to the rehabilitated exercise of urination function for patients with prostate hyperplasia and malignant tumor after prostate surgery. It is urgent to build a scheme of rehabilitated exercise of urination function for postoperative patients of prostate, which based on the fast-track surgery theory and used the time as axis.

The unilateral biportal endoscopic technique is defined as posterior unilateral of two small incision with established percutaneous observation channel and endoscopic observation channel in monitoring field. The operation tools were placed within the channel for each operation inside and outside vertebral canal. The characteristics of dual channel intervention make this technology different from coaxial endoscopic technology. This technique has a clear vision for flexible and convenient operation with the advantages of relatively simple surgical instrument requirements. In recent years, the use of unilateral dual channel endoscopic surgery in treating spinal diseases was gradually increasing, especially in the removal of nucleus including lumbar intervertebral disc protrusion and lumbar spinal stenosis disease decompression, and in endoscopic space-occupying lesions such as vertebral fusion and spinal canal between inside and outside. It was also applied in cervical and thoracic diseases with good clinical effects. Its advantages include less surgical trauma and faster recovery process. In addition, it has a large surgical field of vision under the microscope for easy identification of structures and surgery and with relatively gentle learning curve. Thus, it is conducive to beginners' mastery. However, complications such as nerve injury and postoperative lower limb numbness are still frequently reported. Some factors have to be considered, including insufficient understanding of the new technology, the local anatomy of the dual-channel endoscope, the operation of the spinal canal under the microscope. The successful application of the technique in treating various lumbar diseases needs understanding the operation process of the technique and local anatomy under the microscope.

Objective:To develop a unilateral biportal endoscopic technique and investigate the clinical efficacy of unilateral biportal endoscopic technique in the treatment of lumbar disc herniation and lumbar spinal stenosis.Methods:Between July 2018 and June 2019, 60 patients with lumbar disc herniation or spinal stenosis were treated by unilateral biportal endoscopic technique. According to the inclusion and exclusion criteria, 51 patients were included in the study. There were 25 cases of prolapsed lumbar disc herniation, 26 cases of degenerative lumbar spinal stenosis, all of which were treated with posterior lumbar decompression by unilateral biportal endoscopic laminectomy, and contralateral stealth decompression by unilateral approach was performed on the patients with bilateral stenosis. Endoscopic interbody fusion and percutaneous pedicle screw fixation were performed in 5 patients with instability. Operation time, length of incision, hospital stay and complications were recorded. Visual analogue scale (VAS) for low back pain and leg pain, Oswestry dysfunction index (ODI) and the modified Macnab scale were used to evaluate the clinical efficacy.Results:All operations were completed successfully, and no cases were transferred to open surgery. The operative time was 70.29±19.55 min (44-151 min), while the length of incision was 1.79±0.34 cm (1.4-3.0 cm). Postoperative CT suggested complete decompression with intact contralateral structure. All patients got out of bed 1-3 d after surgery, and the postoperative hospital stay was 3.49±2.76 d (1-14 d). The postoperative follow-up time was 13.59±2.80 months (10-21 months). Forty-six patients returned to work or normal activities within 3 weeks and 5 patients with interbody fusion returned to normal activities within 4 weeks. According to the modified Macnab criteria, the final outcome was excellent in 43 cases, good in 6 cases, and fair in 2 cases. There were 3 cases of dural sac tear during operation and 2 cases of transient numbness of lower limbs after surgery and they all recovered after conservative treatment. The VAS score of low back pain of 26 patients with lumbar spinal stenosis was reduced from 6.69±1.44 before surgery to 3.27±1.43 at postoperative 1 month, 2.69±1.57 at postoperative 3 months, 2.31±1.16 at postoperative 6 months and 2.23±1.28 at the last follow-up, respectively, and the difference was statistically significant ( F=128.534, P<0.005). The VAS scoreof leg pain was reduced from 6.77±1.34 before surgery to 3.27±1.37 at postoperative 1 month, 2.88±1.48 at postoperative 3 months, 2.85±1.52 at postoperative 6 months and 2.54±1.53 at the last follow-up, and the difference was statistically significant ( F=146.951, P<0.005). The ODI score was reduced from 64.18%±8.23% before surgery to 37.53%±4.45% at postoperative 1 month, 27.51%±3.83% at postoperative 3 months, 19.91%±5.27% at postoperative 6 months and 6.84%±2.74% at the last follow-up, and the difference was statistically significant ( F=783.966, P<0.005). The VAS score of low back pain of 25 patients with lumbar disc herniation was reduced from 5.60±1.38 before surgery to 3.04±1.54 at postoperative 1 month, 2.84±1.75 at postoperative 3 months, 3.12±1.86 at postoperative 6 months and 3.44±1.69 at the last follow-up, respectively, and the difference was statistically significant ( F=22.357, P<0.005). The VAS scoreof leg pain was reduced from 5.48±1.45 before surgery to 2.88±1.64 at postoperative 1 month, 2.52±1.83 at postoperative 3 months, 2.76±1.83 at postoperative 6 months and 3.00±1.92 at the last follow-up, and the difference was statistically significant ( F=29.445, P<0.005). The ODI score was reduced from 53.59%±6.87% before surgery to 32.46%±3.78% at postoperative 1 month, 23.39%±2.78% at postoperative 3 months, 16.49%±3.49% at postoperative 6 months and 7.23%±3.15% at the last follow-up, and the difference was statistically significant ( F=790.985, P<0.005). Conclusion:Unilateral biportal endoscopic technique has the advantages of clear and wide field of vision, large operating space, relatively simple surgical instrument need and convenient and flexible operation procedure. It has excellent clinical effects in the treatment of lumbar disc herniation and lumbar spinal stenosis.

Objective:To observe the expression of glucose regulated protein 78 (GRP78) in glioma and its clinical value in evaluating clinical prognosis.Methods:A total of 163 patients with glioma who were surgically removed and confirmed after surgery in the People′s Hospital of Inner Mongolia Autonomous Region were collected from March 2012 to October 2015. Immunohistochemical staining was used to observe the expression of GRP78 in patients with different WHO grades of glioma. Chi square test was used to compare the expression of GRP78 in glioma patients with different age, gender and WHO grade. Whether COX regression analysis GRP78 can be used as an independent prognostic indicator was investigated. Time series test and Kaplan Meier analysis were used to analyze the survival time of patients with different GRP78 expression levels.Results:The expression of GRP78 was up-regulated in glioma, and the patients with high expression of GRP78 (positive expression/strongly positive expression) were more common in stage Ⅲ/Ⅳ than in stage Ⅰ/Ⅱ. The expression of GRP78 was not statistically significant in different age and gender groups ( P>0.05), but was statistically significant in different WHO stages ( P<0.01). GRP78 was an independent prognostic factor for glioma ( P = 0.045). The median survival time of patients with glioma was 28 months (95% CI 37.594 to 47.046), of which the survival time of patients with high expression of GRP78 was 15 months (95% CI 12.922 to 40.801); the survival time of patients with low expression of GRP78 was 35 months (95% CI 39.807 to 51.352), and the difference was statistically significant ( P<0.01). The survival time of patients with high expression of GRP78 was significantly shorter than that of patients with low expression of GRP78 ( χ2 = 13.588, P<0.01). Conclusions:The expression level of GRP78 in glioma is significantly increased; high expression of GRP78 is more common in Ⅲ/Ⅳ glioma, and GRP78 can be used as an effective index to evaluate the poor prognosis of glioma.

Objective: To explore the current status and influencing factors of eating behaviors in adolescents, so as to provide a theoretical foundation for health promotion education among adolescents. Methods: Based on the database from Survey of Chinese Family Health Index (2021), by a random number table method, 1 065 teenagers were selected from the provincial capitals of 22 provinces and 5 autonomous regions in China, as well as 4 municipalities directly under the central government. A general characteristic questionnaire, Patient Health Questionnaire-9 (PHQ-9), Short Form of the Family Health Scale (FHS-SF), 10-item Short Version of the Big Five Personality(BFP-10), Content-based Media Exposure Scale (CM-E) and Sakata Eating Behavior Scale Short Form(EBS-SF) were used to collect information. Multivariate stepwise linear regression analysis was employed to identify and analyze related factors of eating behaviors among adolescents. Receiver operating characteristic was used to validate the predictive ability of the EBS-SF score for overweight and obesity among adolescents. Results: The average scores of BFI-10,C-ME, FHS-SF, PHQ-9 and EBS-SF were (33.08±4.64)(19.20±4.55)(38.48±6.65)(6.09±5.63)(16.75±4.36), respectively. Multivariate linear regression showed that family type (other types), agreeableness, conscientiousness, family health and depression were the main related factors of EBS-SF scores among adolescents( B =2.61,-0.42,0.20,-0.11,0.23, P <0.05).The analysis of receiver operating characteristic curve showed that the EBS-SF scores had a good ability in predicting obesity among male adolescents ( AUC= 0.73, P <0.01). Conclusions Family type, big five personality, family health,depression are the related factors of eating behaviors among adolescents. EBS-SF scores are predictive of obesity in adolescents, which would provide a new perspective for promoting healthy eating habits among adolescents.

Objective: The objective of this study was to verify the lipid-lowering effect of Juhe Fang extract (JHFE) and to determine its characteristic chemical profile in vitro and in vivo. Methods: A hyperlipidemia model was established by feeding mice a high-fat diet (HFD). After treatment for 30 days, serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) levels were measured with an automatic biochemistry analyzer. The components from JHFE obtained from in vivo and in vitro experiments were investigated using an UPLC-Q Exactive-Orbitrap MS/MS. Results: The TC, TG, and LDL-C in the serum significantly decreased and the HDL-C significantly increased after JHFE treatment. A total of 95 compounds from JHEF including 15 phenolic acids (PA), 4 phenyl-ethanoid glycosides (PG), 24 flavonoids (F), 14 triterpenoids (T), 10 diterpenoid glycosides (D), 18 alka-loids (A) and 10 others (O) were identified. Trigonelline was discovered for the first time in a herbal medicine of Juhe Fang. Furthermore, 68 compounds were identified in vivo including 28 prototype compounds and 40 metabolites. The metabolic characteristics of these components were revealed including identification of new metabolites of 4-hydroxyphenyl ethyl-8-O-[α-L- arabinopyranosyl-(1→6)]-β-D-glucopyranoside (PEG) and lirinidine. A total of 43 components from JHFE were absorbed and/or metabolized. The contribution rate of each type of chemical component from JHFE to its lipid-lowering effect from high to low were A, F, PG, PA, D and T. Conclusion: The results of this study showed that JHFE demonstrated a significant lipid-lowering effect in a high-fat diet (HFD)-induced hyperlipidemia mouse model. Specific types of PA, PG, F, D, T and A formed the pharmaceutical architecture of the lipid-lowering effect of JHFE. This study should prove useful for clarifying the components responsible for the lipid-lowering effect of JHFE and provide a basis for precision quality control research.