Ginsenoside Rg1 relieves rat intervertebral disc degeneration and inhibits IL-1ß-induced nucleus pulposus cell apoptosis and inflammation via NF-κB signaling pathway.

The study aimed to investigate the effect of ginsenoside Rg1 on intervertebral disc degeneration (IVDD) in rats and IL-1ß-induced nucleus pulposus (NP) cells, and explore its underlying mechanism. Forty IVDD rat models were divided into the IVDD group, low-dose (L-Rg1) group (intraperitoneal injection of 20 mg/kg/d ginsenoside Rg1), medium-dose (M-Rg1) group (intraperitoneal injection of 40 mg/kg/d ginsenoside Rg1), and high-dose (H-Rg1) group (intraperitoneal injection of 80 mg/kg/d ginsenoside Rg1). The pathological change was observed by HE and safranin O-fast green staining. The expression of IL-1ß, IL-6, TNF-α, MMP3, aggrecan, and collagen II was detected. The expression of NF-κB p65 in IVD tissues was detected. Rat NP cells were induced by IL-1ß to simulate IVDD environment and divided into the control group, IL-1ß group, and 20, 50, and 100 µmol/L Rg1 groups. The cell proliferation activity, the apoptosis, and the expression of IL-6, TNF-α, MMP3, aggrecan, collagen II, and NF-κB pathway-related protein were detected. In IVDD rats, ginsenoside Rg1 improved the pathology of IVD tissues; suppressed the expression of IL-1ß, IL-6, TNF-α, aggrecan, and collagen II; and inhibited the expression of p-p65/p65 and nuclear translocation of p65, to alleviate the IVDD progression. In the IL-1ß-induced NP cells, ginsenoside Rg1 also improved the cell proliferation and inhibited the apoptosis and the expression of IL-6, TNF-α, aggrecan, collagen II, p-p65/p65, and IκK in a dose-dependent manner. Ginsenoside Rg1 alleviated IVDD in rats and inhibited apoptosis, inflammatory response, and ECM degradation in IL-1ß-induced NP cells. And Rg1 may exert its effect via inhibiting the activation of NF-κB signaling pathway.

Sex difference for the risk of amputation in diabetic patients: A systematic review and meta-analysis.

The risk of amputation is a sequelae of diabetic foot ulceration, which are significantly increased in diabetic patients and caused huge morbidly and mortality. However, whether the risk amputation in diabetic patients are differing in male and female remains inconclusive. We therefore conducted a systematic review and meta-analysis to assess the sex difference for the risk of amputation in diabetic patients. We systematically searched PubMed, EmBase, and the Cochrane library to identify eligible study from their inception up to November 2020. The diagnostic value of male patients on subsequent amputation risk were assessed by using sensitivity, specificity, positive and negative likelihood ratio (PLR and NLR), diagnostic odds ratio (DOR), and area under the receiver operating characteristic curve (AUC). Twenty-two studies recruited a total of 33,686,171 diabetic patients were selected for quantitative analysis. The risk of amputation in male diabetic patients was greater than female diabetic patients (DOR: 1.38; 95%CI: 1.13-1.70; P<0.001). The sensitivity and specificity for male diabetic patients on the risk of amputation were 0.72 (95%CI: 0.72-0.73), and 0.51 (95%CI: 0.51-0.51), respectively. Moreover, the PLR and NLR of male diabetic patients for predicting amputation were 1.13 (95%CI: 1.05-1.22), and 0.82 (0.72-0.94), respectively. Furthermore, the AUC for male diabetic patients on amputation risk was 0.56 (95%CI: 0.48-0.63). This study found male diabetic patients was associated with an increased risk of amputation than female diabetic patients, and the predictive value of sex difference on amputation risk in diabetic patients was mild.

Circular RNA_CNST Promotes the Tumorigenesis of Osteosarcoma Cells by Sponging miR-421.

Circular RNAs (circRNAs) act crucial roles in the progression of multiple malignancies including osteosarcoma (OS). But, the underlying mechanisms by which hsa_circ_0017311 (circCNST) contributes to the tumorigenesis of OS remain poorly understood. Our present study aimed to explore the role and mechanisms of circCNST in OS tumorigenesis. The differentially expressed circRNAs were identified by the Gene Expression Omnibus database. The association of circCNST with clinicopathological features and prognosis in patients with OS was analyzed by RNA fluorescence in situ hybridization (FISH) and quantitative real-time polymerase chain reaction (PCR) analysis. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), colony formation assays, and a xenograft tumor model were conducted to assess the role of circCNST in OS cells in vitro and in vivo. CircCNST-specific binding with miR-421 was confirmed by FISH, luciferase gene report, and RNA immunoprecipitation assays. As a result, we found that the expression levels of circCNST were dramatically increased in OS tissues and cell lines as compared with the adjacent normal tissues, and it was associated with tumor size and poor survival in OS patients. Knockdown of circCNST repressed cell viability, colony formation, and xenograft tumor growth, while restored expression of circCNST reversed these effects. Furthermore, circCNST was colocalized with miR-421 in the cytoplasm and acted as a sponge of miR-421, which attenuated circCNST-induced proliferation-promoting effects in OS cells by targeting SLC25A3. In conclusion, our findings demonstrate that circCNST promotes the tumorigenesis of OS cells by sponging miR-421, and provides a potential biomarker for patients with OS.

Astragalus polysaccharide ameliorates lipopolysaccharide-induced cell injury in ATDC5 cells via miR-92a/KLF4 mediation.

BACKGROUND: Astragalus polysaccharide (APS) is a traditional Chinese herbal medicine with anti-inflammatory and anti-aging activities. OBJECTIVE: This study aimed to explore the effect and associated mechanisms of APS on LPS-induced injury in ATDC5 cells, to evaluate the potential of APS for use as an adjuvant therapy for osteoarthritis (OA). MATERIALS AND METHODS: ATDC5 cells were pre-treated with APS and stimulated with lipopolysaccharide (LPS). Cell viability, ROS generation as well as the expression of IL-6, TNF-α, iNOS and Cox-2 were evaluated by performing CCK8 assay, ROS detection, ELISA, western blot and qRT-PCR. The expression of NF-κB and p38MAPK signal pathways related proteins and KLF4 was measured through western blot. RESULTS: LPS increased the expression of IL-6 and TNF-α, elevated the expression of Cox-2, iNOS and increased ROS generation. APS treatment significantly alleviated LPS-induced damage in ATDC5 cells. Besides, miR-92a was down-regulated while KLF4 was up-regulated by APS. At the same time, the targeting relationship between miR-92a and KLF4 was demonstrated. The inhibitory effects of APS on LPS-induced injury in ATDC5 cells were attenuated by the combination of KLF4 siRNA. In addition, LPS induced NF-κB and p38MAPK signal pathways were decreased by APS treatment. Also, the inhibitory effect of APS on NF-κB and p38MAPK signal pathways was reversed by KLF4 siRNA. CONCLUSIONS: The present study reveals that APS protects ATDC5 cells against LPS induced-injury by regulation of miR-92a/KLF4 axis and suppressing NF-κB and p38MAPK signal pathways.

SLC25A22 Promotes Proliferation and Metastasis of Osteosarcoma Cells via the PTEN Signaling Pathway.

Osteosarcoma is a highly malignant bone tumor. However, due to the high complexity of the occurrence and metastasis of osteosarcoma, the exact mechanism promoting its development and progression remains to be elucidated. This study highlights the causal link between solute carrier family 25 member 22 (SLC25A22) and the development, progression, and metastasis of osteosarcoma. SLC25A22 is upregulated in human osteosarcoma and predicts a poor prognosis. The upregulation of SLC25A22 in osteosarcoma tissues was significantly associated with cell proliferation, invasion, and metastasis. Studies of functional gain (overexpression) and loss (knockdown) showed that SLC25A22 significantly increases the ability of osteosarcoma cells to proliferate, as well as invade and metastasize in vitro. At the same time, the expression of SLC25A22 promoted the progression of the cellcycle of osteosarcoma cell lines and inhibited the apoptosis of osteosarcoma cells. Analysis using a mouse xenograft model showed that xenografts of SLC25A22 stable overexpressing osteosarcoma cells had a significant increase in tumor volume and weight compared to the control group. Lung metastasis models in mice showed that expression of SLC25A22 promoted lung metastasis of osteosarcoma in vivo. Furthermore, SLC25A22 inhibited phosphatase and tensin homolog expression and increased phosphorylation of protein kinase b (Akt) and Focal Adhesion Kinase (FAK) in the phosphatase and tensin homolog signaling pathway. In summary, SLC25A22 is highly expressed in osteosarcoma, promoting osteosarcoma cell proliferation and invasion by inhibiting the phosphatase and tensin homolog signaling pathway.

[Radiographic analysis of the osseous fixation zone for the iliac crest external fixation with Schanz screws].

OBJECTIVE: To radiographically analyze the osseous fixation zone for the iliac crest external fixation with Schanz screws and in order to guide their placement. METHODS: Nine adults with 2.0-mm-slice continuous pelvic axial CT scans were selected as research subjects. Each CT scan data was imported into MIMICS 10.0. The osseous fixation zone the upper portion of the anterior column of the acetabulum which is located between the anterior superior iliac spine and the gluteal medius pillar and between the iliac crest and the acetabulum-for the iliac crest external fixation with Schanz screws was reconstructed into true sagittal and true coronal planes by using the software. Then the measurements were taken on the reconstructed planes with measuring tools. Finally, the measured data was analyzed. RESULTS: The palpable iliac crest segment, which was of 49.6 mm width and located 16.5 mm posterior to the anterior superior iliac spine could be used to locate the start points of the Schanz screws. Under the above-mentioned iliac crest segment, the osseous zone was deep, got ample bony materials and could intraosseously contain Schanz screws with 5.0 mm diameter. The screws could be safely inserted to a minimal depth of 71.7 mm towards the acetabular dome and to a maximal depth of 143.5 mm posterior to the acetabulum. CONCLUSION: The study can guide the effective insertion of the iliac crest Schanz screws. By setting a suitable start point in the above-mentioned iliac crest region and angling correctly relative to the acetabulum,the Schanz screw can be inserted into the relative strong cancellous bone above or posterior to the acetabulum with a considerable depth, to getting more bone engagement.

[Radiographic anatomical analysis of the pelvic Teepee view].

OBJECTIVES: To research radiographic anatomy of the main structure of the pelvic Teepee view, including its azimuth direction and view anatomy structure. METHODS: From June 2013 to June 2014 adult pelvic CT examination results were filtered, excluding skeletal deformities and pelvic osseous destruction caused by tumors, trauma, etc. The data of 2.0 mm contiguous CT scan of 9 adults' intact pelves was,selected and input into Mimics 10.01 involving 7 males and 2 females with an average age of (41.2±10.3) years old. Utilizing the software, the 3D CT reconstructions of the pelves were completed. Setting the transparency being high,the pelvic 3D reconstructions were manipulated from the pelvic anteroposterior view to the combined obturator oblique outlet view and fine-tuned till the regular Teepee-or teardrop-shaped appearance emerges. Cutting tools of the software were at the moment applied to separate the "Teepee" from the main pelvis for each reconstruction. Then the "Teepee" and the rest (main) part of the pelvis were displayed in different color to facilitate the analysis on the Teepee, iliac-oblique, and anteroposterior views. RESULTS: The "Teepee" started from the posterolateral aspect of the anterior inferior iliac spine and finished at the cortex between the posterior superior iliac spine and the posterior inferior iliac spine in a direction of being from caudal-anterior-lateral to cranial-posterior-medial. The radiographic anatomical composition of the "Teepee" contained one tip, one base,and two aspects. With the inner and outer iliac tables being the inner and outer aspects of the "Teepee", the tip is consequently formed by their intersection. The base is imaged from the cortex of the greater sciatic notch. The medial-inferior-posterior portion of the "Teepee" contains a small part of sacroiliac joint and its corresponding side of bone of the sacrum. CONCLUSIONS: The "Teepee" is a zone of ample osseous structures of the pelvis, aside from a small medial-inferior-posterior portion, the main zone of which can be accepted as a safe osseous zone for the anchor of implants stabilizing certain pelvic and acetabular fracture patterns. The Teepee view can be utilized as guidance for the safe percutaneous insertion of such implants.

[S2 iliosacral screw insertion technique].

OBJECTIVE: To introduce a technique pertaining to S2 iliosacral screw insertion. METHODS: The screw pathway was first measured on the preoperative pelvic CT scan or the standard sacral lateral radiograph to make sure the existence of the "safe zone" in the S2 segment for screw insertion. Under general anesthesia, patients were positioned supine or prone, depending on the injury pattern of pelvic ring or associated injuries requiring concomitant operation. The operation field was routinely sterilized using iodine and subsequent alcohol solution and draped. The tip of a guide wire was inserted through a stab wound to the posterior outer iliac table, manipulated in the "safe zone" being enclosed by the anterior aspect of the S2 nerve root tunnel, the anterior aspect of the sacral vertebrae, and the inferior aspect of the S1 foramen under the guidance of the standard sacral lateral fluoroscopy, and then the tip was hammered one to two millimeters into the iliac cortex. The guide wire progressed along the trajectory between the inferior aspect of the S1 foramen and the superior aspect of the S2 foramen on the pelvic outlet fluoroscopic view, and then along the posterior to the anterior aspect of the S2 sacral vertebrae and alae on the pelvic inlet fluoroscopic view with a predetermined length. At that moment, in order to ensure the safety, another standard sacral lateral view was imaged to detect the guide wire's tip which should locate posterior to the anterior aspect of the sacral vertebrae and anterior to the anterior aspect of the S2 nerve root tunnel. Subsequently, the depth was measured, the trajectory was drilled and tapped, and the screw was inserted. Following the removal of the guide wire, the wound was irrigated and sutured. RESULTS: Utilizing this insertion technique, there were 30 S2 iliosacral screws in total being placed to stabilize the injured and unstable posterior pelvic ring in 27 patients. Each S2 screw was accompanied by an ipsilateral S1 screw. The S2 screw location was completely intraosseous in all patients, which was verified by postoperative pelvic outlet and inlet radiographs and CT scans. The insertion accuracy was 100 percent in the present series. CONCLUSION: The S2 iliosacral screw insertion technique is safe and reproducible to guide the placement of the S2 screw, enhancing the stability for the compromised posterior pelvic ring.

[Influence of the included angle between anterior aspects of S2 and S1 vertebral bodies on pelvic inlet imaging in mid-line sagittal plane].

OBJECTIVE: To analyze the influence of included angle between the anterior aspects of S2 and S vertebral bodies on pelvic inlet imaging in the pelvic midline sagittal plane. METHODS: Totally 58 axial pelvic CT scans were chosen as study objects including 43 males and 15 females,with an average age of 40.7 years old (ranged,18 to 68 years old). The angles between the anterior aspects of S2 and S1, vertebral bodies and the horizontal plane on midline sagittal CT reconstruction were measured to simulate the optimal S2 and S1 inlet angles. The included angle between the anterior aspects of S2 and S1 vertebral bodies was calculated by subtrocting the S1,inlet angle from the S2 inlet angle defined as a base number. Then, the impact of the calculated included angles on the pelvic inlet imaging was analyzed. Results:The S2 inlet angles averaged (30.5±6.5) degrees; the S inlet angles averaged (25.7±5.9) degrees. The difference between them was significant (t=3.35, P=0.001). Ten patients had zero angle between the anterior aspects of S2 and S1 vertebral bodies; 14 patients had negative angle, averaged-(8.9±8.1) degrees; 34 patients had positive angle,averaged (11.8+6.4) degrees. CONCLUSION: The difference of included angle between the anterior aspects of S2 and S1 vertebral bodies leads to the difference between S1 inlet view and S2 inlet view in most cases, complicating the pelvic inlet imaging,and affecting the reliability of the application of pelvic inlet view. Utilizing the angles measured on the preoperative midlihe sagittal CT reconstruction to obatin the patient-customized S1 and S2 inlet views could accurately guide the S1 and S2 iliosacral screw insertion.

[Effect of the anterior aspect of sacral nerve root tunnel on iliosacral screw placement on the standard lateral image of sacrum].

OBJECTIVE: To introduce the location and course of S1, S2 sacral nerve root tunnel and to clarify the significance of the anterior aspect of sacral nerve root tunnel on placement of iliosacral screw on the standard lateral sacral view. METHODS: Firstly the data of 2.0 mm slice pelvic axial CT images were imported into Mimics 10.0, and the sacrum, innominate bones, and sacral nerve root tunnels were reconstructed into 3D views respectively, which were rotated to the standard lateral sacral views, pelvic outlet and inlet views. Then the location and course of the S1, S2 sacral nerve root tunnel on each view were observed. RESULTS: The sacral nerve root tunnel started from the cranial end and anterior aspect of the vertebral canal of the same segment and ended up to the anterior sacral foramen with a direction from cranial-posterior-medial to caudal-anterior-lateral. The tunnel had a lower density than the iliac cortex and greater sciatic notch on the pelvic X-rays,especially on the standard sacral lateral view, on which it showed up as a disrupted are line and required more careful recognition. CONCLUSION: It can prevent the iliosacral screw from penetrating the sacral nerve root tunnel and vertebral canal when recognizing the anterior aspect of sacral nerve root tunnel and choosing it as the caudal-posterior boundary of the "safe zone" on the standard lateral sacral view.

[Classification of upper sacral segment based on continuous axial pelvic computed tomography scan].

OBJECTIVES: To introduce a classification system of upper sacral segment and its significance based on the continuous pelvic axial computed tomography scan. METHODS: The whole pelvis 2.0 mm thick axial scan images of 127 cases were observed, the sacroiliac screw channel of S1 were measured, according to the size of the transverse screw channel the upper sacral segment were classified. Such as transverse screw channel existed and in at least 4 layer scan images its width was > 7.3 mm, it was defined as sacral segment of the normal type. Such as transverse screw channel existed and its maximum width was 7.3 mm or less on scanning level, it was defined as a transitional. Such as transverse channel did not exist, or its width on all scanning level was 0 mm or less, it was defined as dysplastic. Various cases,percentage, and the average of the transverse screw channel were calculated. RESULTS: There were 58 normal (45.7%),42 transitional (33.1%), and 27 dysplastic (21.2%) upper sacral segments with an averaged width of the tansverse screw channel of 13.9 mm, 5.2 mm, and 0.9 mm, respectively. Each specimen could be defined as one of the three types of upper sacral segment without exceptions. CONCLUSION: It is possible to insert a transverse iliosacral screw into a normal upper sacral segment when indicated because of the capacious transverse screw channel. The transverse iliosacral screw placement into the transitional and dysplastic upper sacral segments was contraindicated because of the limited or none transverse screw channel. The transitional upper sacral segment was superior to the dysplastic segment due to its starting point location restriction on the true lateral sacral view.

Analysis of long-term outcomes of double-strut bone graft for osteonecrosis of the femoral head.

OBJECTIVE: To analyze the long-term effect of double-strut bone graft for osteonecrosis of the femoral head (ONFH). METHODS: A total of 366 adult patients with ONFH in 466 hips underwent double-strut bone graft from March 1988 to January 1999. Of them, 186 patients with 206 hips and an average age of 32.2 years (range, 20-60 years) were followed up for more than five years, up to January 2006. Based on the Association Research Circulation Osseous (ARCO) classification, there were 36 hips in stage IIB, and 30, 40, 40, 32 and 28 in stage IIC, IIIA, IIIB, IIIC and IV, respectively. The functional results of affected hips were evaluated by the hundred forked method. RESULTS: Hip pain in all patients disappeared or alleviated greatly after the operation. The height of the femoral head improved to various extents, and the range of motion of the hip joint increased. The patients were followed up for 5-16 years, (average 10.5 years). The total scores increased significantly postoperatively (P < 0.01). The rate of excellent and good results was 83.3%, 80.0%, 75.0%, 65.0%, 40.6% and 28.6% in stage IIB, IIC, IIIA, IIIB, IIIC and IV, respectively (63.6% for the whole group). CONCLUSION: The long-term effect of double-strut bone graft for ONFH is satisfactory in relation to staging of ONFH. Favorable results can be expected in young ONFH patients in stage IIB, IIC, and IIIA, IIIB.