[Retracted] SOX2 knockdown inhibits the migration and invasion of basal cell carcinoma cells by targeting the SRPK1­mediated PI3K/AKT signaling pathway.

[This retracts the article DOI: 10.3892/ol.2018.9810.].

Metformin promotes angiogenesis and functional recovery in aged mice after spinal cord injury by adenosine monophosphate-activated protein kinase/endothelial nitric oxide synthase pathway.

Treatment with metformin can lead to the recovery of pleiotropic biological activities after spinal cord injury. However, its effect on spinal cord injury in aged mice remains unclear. Considering the essential role of angiogenesis during the regeneration process, we hypothesized that metformin activates the adenosine monophosphate-activated protein kinase/endothelial nitric oxide synthase pathway in endothelial cells, thereby promoting microvascular regeneration in aged mice after spinal cord injury. In this study, we established young and aged mouse models of contusive spinal cord injury using a modified Allen method. We found that aging hindered the recovery of neurological function and the formation of blood vessels in the spinal cord. Treatment with metformin promoted spinal cord microvascular endothelial cell migration and blood vessel formation in vitro. Furthermore, intraperitoneal injection of metformin in an in vivo model promoted endothelial cell proliferation and increased the density of new blood vessels in the spinal cord, thereby improving neurological function. The role of metformin was reversed by compound C, an adenosine monophosphate-activated protein kinase inhibitor, both in vivo and in vitro, suggesting that the adenosine monophosphate-activated protein kinase/endothelial nitric oxide synthase pathway likely regulates metformin-mediated angiogenesis after spinal cord injury. These findings suggest that metformin promotes vascular regeneration in the injured spinal cord by activating the adenosine monophosphate-activated protein kinase/endothelial nitric oxide synthase pathway, thereby improving the neurological function of aged mice after spinal cord injury.

Utx Regulates the NF-κB Signaling Pathway of Natural Stem Cells to Modulate Macrophage Migration during Spinal Cord Injury.

Neural stem cells (NSCs) play vital roles in the homeostasis of neurological function. Ubiquitously transcribed tetratricopeptide repeat, X chromosome (UTX) is an important regulator of stem cell phenotypes. In our current study, we aimed to investigate whether the conditional knockout of UTX on neural stem cells alters macrophage assembly in response to spinal cord injury (SCI). Conditional knockout Utx of NSC (Utx-KO) mice was used to generate SCI models by the modified Allen method. We reported that neurological function and scar hyperplasia significantly improved in Utx-KO mice after SCI, accompanied by significantly reduced assembly of macrophages. With a 45-fold pathway array and Western blot, we found that Utx-KO could significantly inhibit NF-κB signaling activation and promote the synthesis and secretion of macrophage migration inhibitory factor (MIF) in NSCs. Administration of the selective NF-κB p65 activator betulinic acid and the selective MIF inhibitor ISO-1 confirmed that the activation of NF-κB p65 phosphorylation or inhibition of MIF could eliminate the benefits of Utx-KO in SCI, such as inhibition of macrophage aggregation and reduction in scar proliferation. This study confirmed that UTX in NSCs could alter macrophage migration and improve neurological function recovery after SCI in mice.

[Study on protective effect of water extract from Sabia parviflora on liver injury in mice induced by acetaminophen].

The aim of this study was to observe the protective effect of water extract from Sabia parviflora on mice with acute liver injury induced by acetaminophen, and investigate its possible mechanism. Fifty-eight Kunming mice were divided into 6 groups, 8 in the normal group, 10 in the model group, 10 in the biphenyl diester group, and 10 each in the low, medium and high dose groups. After adaptive feeding for one week, the mice in normal group were intragastrically administered with an equal volume of 0.5% sodium carboxymethylcellulose sodium(CMC-Na), and the mice in other groups were intragastrically administered with corresponding drugs at 20 mL·kg~(-1) once a day. Then acetaminophen(200 mg·kg~(-1)) was administered after the above drug administration except the normal group. The behavior and signs of the experimental animals were observed every day and the samples were taken for experiments on the next day of the final administration. The liver mass and mass index were calculated. The blood was collected from the abdominal aorta and centrifuged to obtain the serum for detecting aspartate aminotransferase(AST) activity and alanine aminotransferase(ALT) activity. The liver tissue homogenate was used to detect superoxide dismutase(SOD) activity, glutathione(glutathione, r-glutamyl cysteingl+glycine, GSH) activity and malondialdehyde(MDA) content. Liver tissue was analyzed for histological analysis. The results showed that S. parviflora could alleviate the lipid peroxidation damage in the liver caused by acetaminophen, reduce the ALT and AST activities in serum, increase the levels of SOD and GSH in liver tissue, decrease the content of MDA in liver tissue, and inhibit the apoptosis. S. parviflora could also improve the live histopathological profile, protect liver cells and restore liver function. Among them, the high dose had the most significant effect and showed dose-effect relationship. This study indicated that S. parviflora had a significant protective effect on acetaminophen-induced liver injury in mice, and its mechanism may be related to its anti-oxidation effect and inhi-bitory effect on apoptosis.

SOX2 knockdown inhibits the migration and invasion of basal cell carcinoma cells by targeting the SRPK1-mediated PI3K/AKT signaling pathway.

Basal cell carcinoma (BCC) is the most common type of human skin cancer, which is driven by the aberrant activation of Hedgehog signaling. Previous evidence indicated that sex determining region Y-box 2 (SOX2) is associated with the tumor metastasis. However, the expression and role of SOX2 in BCC remain unknown. Therefore, the aim of the current study was to analyze the possible mechanism of SOX2 in the progression of BCC. The levels of SOX2 in BCC cells were detected by reverse transcription-quantitative polymerase chain reaction. Transwell assays were also used to determine the migration and invasion of BCC cells. Immunoblotting and immunofluorescence were used for analyzing the role of SOX2 knockdown in the serine-arginine protein kinase 1 (SRPK1)-mediated phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) signaling pathway in BCC cells. The results demonstrated that SOX2 is overexpressed in BCC tissues and cells. In addition, SOX2 knockdown inhibited the migration and invasion of BCC cells, and the epithelial-mesenchymal transition (EMT) progress of BCC cells. It was also observed that SOX2 knockdown decreased SRPK1 expression, which further led to the downregulation of PI3K and AKT expression levels in BCC cells. Furthermore, SRPK1 transfection or PI3K/AKT pathway activation abolished the inhibitory effects of SOX2 knockdown on the migration, invasion and EMT progress of BCC cells. In conclusion, these results indicated that SOX2 may potentially serve as a target for BCC therapy by targeting the SRPK1-mediated PI3K/AKT signaling pathway.

Tetramethylpyrazine Facilitates Functional Recovery after Spinal Cord Injury by Inhibiting MMP2, MMP9, and Vascular Endothelial Cell Apoptosis.

BACKGROUND: Spinal cord injury (SCI) is a major public health issue that leads to neurological dysfunctions and morbidities in patients. Tetramethylpyrazine (TMP) plays a neuroprotective role in SCI; however, the underlying mechanism has not been fully elucidated. OBJECTIVE: In the present study, we aimed to investigate the mechanisms and therapeutic effects of TMP on SCI. METHODS: A contusion SCI model was established that used a modified Allen's method. In the TMP group, TMP (200 mg/kg) was injected daily for 5 days post-injury, while in the Negative Control (NC) group, an equal volume of normal saline was injected. Hindlimb motor function was evaluated using the Basso, Beattie, Bresnahan (BBB) scale. The effects of TMP on protein levels of the matrix metalloproteinases 2 (MMP2) and 9 (MMP9), Bax and cleaved caspase-3 were determined by western blotting. Apoptotic changes in vascular endothelial cells were evaluated using immunofluorescence and TUNEL staining. Alterations in 3D vessel morphology after treatment with TMP were assessed by synchrotron radiation micro-CT (SRµCT). RESULTS: TMP treatment significantly improved recovery in hindlimb motor function and attenuated vascular endothelial cell apoptosis in rats with SCI. Additionally, TMP treatment markedly decreased the protein levels of MMP2 and MMP9, pro-apoptotic bax and cleaved caspase-3 while promoting angiogenesis, as evidenced by vessel visualization using SRµCT. CONCLUSION: These results indicate that TMP attenuated SCI-induced neurological impairments by the down-regulation of the expression of MMP2 and MMP9 proteins, the inhibition of vascular endothelial cell apoptosis, and the promotion of angiogenesis.

Modified O-T advancement flap for reconstruction of skin defects.

BACKGROUND: Round or oval defects are common in skin surgery. Functional and cosmetical reconstruction of defects in reparative process is critical for patients. Various flaps have been described, however, these flaps often result in longer scar or tip necrosis. To overcome these shortcomings, we modified O-T advancement flap on the basis of conventional O-T flap and observed the validity and complications during defect closure. MATERIALS AND METHODS: Defect transverse diameter was marked along the direction of minimum tension at the circular center. Extended line was drawn along defect transverse diameter with the same length of circular diameter. The skin was cut apart, and the flap was separated under the skin. Then the flap tips were sutured and fixed with the opposite center. After drainage, the defects were bandaged under compression. RESULTS: This study includes a total number of 48 patients. We examined the location and size of defect and postoperative clinical courses. The follow-up period was from 3 months to 1 year. Overall, 41 of 48 patients achieved the satisfactory postoperative effect. Recurrence and limb dysfunction complication was not observed, except 2 cases of wound scar, 3 cases of wound infect and 2 cases of flap tip necrosis. CONCLUSION: Modified O-T advancement flap is practical and safety. It overcomes the shortcomings of traditional O-T flap. Reconstruction of modified O-T flap is aesthetically acceptable.

Synchrotron radiation micro-CT as a novel tool to evaluate the effect of agomir-210 in a rat spinal cord injury model.

MicroRNA-210 (miR-210) was initially reported to be associated with hypoxia and plays a vital role in modulating angiogenesis. However, the potential effect and underlying mechanisms of miR-210 activity in rat spinal cord injury (SCI) have not yet been fully illuminated. In the present study, differential microRNA expression after SCI was determined by Microarray analysis. To explore the effect of miR-210 after SCI, we intrathecally injected agomir-210 with Alzet Osmotic Pumps to up-regulated the endogenous miR-210 expression. Then, synchrotron radiation micro-CT (SRµCT) imaging was used to investigate the effect of agomir-210 in rat SCI model. We found that the endogenous miR-210 expression could be up-regulated by intrathecal agomir-210 injection. The administration of agomir-210 significantly promoted angiogenesis, as evidenced by increased vessel number and volume detected by SRµCT, attenuated the lesion size and improved functional recovery after SCI. Additionally, agomir-210 attenuated cellular apoptosis and inflammation in the injured rat spinal cord. Expression levels of pro-apoptotic protein (Bax) and pro-inflammatory cytokines (TNF-α and IL-1ß) were significantly decreased after agomir-210 treatment, whereas expression levels of anti-apoptotic (Bcl-2) and anti-inflammatory (IL-10) proteins were up-regulated. In conclusion, our results indicated that SRµCT is a powerful imaging tool to evaluate the effects of angiogenesis after agomir-210 administration in rat SCI model. The up-regulation of endogenous miR-210 expression following agomir-210 administration promoted angiogenesis and anti-apoptotic protein expression, and attenuated inflammation. MiR-210 played a positive role in neurological functional recovery and could be a potential new therapeutic target for SCI.

Tetramethylpyrazine enhances functional recovery after contusion spinal cord injury by modulation of MicroRNA-21, FasL, PDCD4 and PTEN expression.

Our previous study showed Tetramethylpyrazine (TMP) has protective effects against SCI. In this study, we aimed to uncover the mechanism underlying the protective effects of TMP in SCI. SCI was induced in Sprague-Dawley rats with a modified weight-drop device. One group was subjected to SCI in combination with TMP administration at a dose of 200mg/kgd, for 3 days. Concurrently, another group received SCI in combination with an equal volume of 0.9% saline. Locomotor functional recovery was assessed during the 4 weeks post-injury by performing the Basso, Beattie, and Bresnahan (BBB) rating procedure. Lesion size and spared tissue were measured by cresyl violet staining. MicroRNA-21 (miR-21) expression was determined by real-time PCR and in situ hybridization. FasL, PDCD4, and PTEN are direct targets of miR-21 in many diseases and cell types; their levels were analyzed by western blot. Immunohistochemistry was performed to observe the expression of PDCD4 and PTEN. Cell apoptosis was assessed by TUNEL staining and DNA laddering. TMP treatment after contusion SCI significantly improved functional recovery, decreased lesion size, and increased tissue sparing and miR-21 levels; expression of FasL, PDCD4, and PTEN was decreased. TMP treatment also reduced apoptosis after SCI. Thus, TMP administration improved functional recovery and reduced cell apoptosis. Its protective effect may partly based on increasing the expression of miR-21 and decreasing the expression of FasL, PDCD4, and PTEN. These could serve as new exploratory targets for SCI treatment.

New isoindolinones from the fruiting bodies of Hericium erinaceum.

Hericium erinaceus is a well-known medicinal and edible mushroom, which is considered as a potential source to obtain antitumor candidates. In this work, five new isoindolinones, named erinaceolactams A-E (1-5), along with five known compounds (6-10), were isolated from 70% ethanol extract of the fruiting bodies of H. erinaceus. The structures of new compounds were validated by HRESIMS and 1D, 2D NMR. It's worth mentioning that there are two pairs of isomers included in the new compounds. Moreover, their cytotoxicity against metastatic human hepatocellular carcinoma cell lines SMMC-7221 and MHCC-97H were evaluated. The results showed that compounds 6 and 7 exhibited promising inhibitory potency against the growth of two cell lines.

Excision of apocrine glands with preservation of axillary superficial fascia for the treatment of axillary bromhidrosis.

BACKGROUND: Axillary bromhidrosis is a distressing problem, which has a strong negative effect on one's social life. OBJECTIVE: To evaluate the effects and complications of the surgical modality for the treatment of axillary bromhidrosis. METHODS: One hundred fifteen patients with axillary bromhidrosis were treated. Two incisions were made transversely along the marked lines on the axillary crease. Subdermal undermining of the marked area with a depth of 0.3 to 0.5 cm and transverse detachment were performed, allowing the exposure of the skin flaps. Skin flaps were carefully separated from the skin. The apocrine glands, follicles, and fats were dissected, and the axillary superficial fascia was maintained. RESULTS: All patients achieved good results in terms of malodor elimination during the follow-up period. All patients reported reduction in axillary sweating. Postoperative complications were minor, including small hematoma (3 cases), delayed wound healing (5 cases), pressure blister (5 cases), and slightly wound scar (2 cases). No infection, skin necrosis, malodor, or recurrence was observed. One hundred eleven patients (96.5%) were very satisfied and 4 (3.5%) patients satisfied with the procedure, with none regretful. CONCLUSION: The procedure has the advantage of a high success rate in radical elimination of the malodor with minor complications.

Anterior cervical discectomy with arthroplasty versus anterior cervical discectomy and fusion for cervical spondylosis.

This meta-analysis aims to estimate the benefits and drawbacks associated with anterior cervical discectomy with arthroplasty (ACDA) versus anterior cervical discectomy and fusion (ACDF) for cervical spondylosis. Of 3651 identified citations, 10 randomised controlled studies involving 2380 participants were included. Moderate quality evidence supports that patients in the ACDA group had: (1) a higher Neck Disability Index (NDI) success rate at 3 month (relative risk [RR]=0.85, 95% confidence interval [CI] 0.78 to 0.93, p=0.0002) and 2 year follow-up (RR=0.95, 95%CI 0.91 to 1.00, p=0.04); (2) greater neurological success at 2 year follow-up (RR=0.95, 95%CI 0.92 to 0.98); and (3) were more likely to be employed within 6 weeks after surgery (RR=0.80 95%CI 0.66 to 0.96). In summary, the current evidence indicates that ACDA is associated with a higher NDI success rate in the short and long-term as well as a higher neurological success rate. Patients who undergo ACDA may also have a greater likelihood of being employed in the short-term. However, all of the evidence reviewed is of moderate or low quality and the clinical significance often marginal or unclear. Additional data are needed to compare the benefits and limitations of ACDA and ACDF.

[Tetramethylpyrazine accelerated spinal cord repair through regulation of caspase-3 and neurofilament protein expression: an acute spinal cord injury model in rats].

OBJECTIVE: To investigate the effect of tetramethylpyrazine on spinal cord repair through hypothesized pathways including regulation of caspase-3, neurofilament protein NF-L, NF-H, and NF-M expression. METHODS: Allenos weight drop method was used to establish acute spinal cord injury (SCI) rat model at T10 section and was conducted in 80 adult Sprague Dawley rats that were subsequently divided into pseudo-therapy group (sodium chloride group, n=40) and tetramethylpyrazine group (treatment group, n=40). Eight normal rats were included into the pseudo-surgery normal group (n=1 for each time point). The function of spinal cord was evaluated with animal behavioral scores by measuring modified Rivilin loxotic plate degree and counting BBB score at 1, 3, 7, 14, and 21 d postoperatively. The injured spinal cord tissue samples were harvested at 1 h, 3 h, 6 h, 1 d, 3 d, 7 d, 14 d, and 21 d postoperatively (n=5 for each time point) for the preparation of continuous histological sections that were analyzed the expression of caspase-3,NF-L,NF-H, and NF-M by immunohistochemistry method. RESULTS: At postoperative Day 7, 14, and 21, animal behavioral scores revealed higher modified Rivlin loxotic plate degree and BBB scores in the treatment group when compared with those of the control group (P<0.05). The cells in the injured spinal cord tissue expressed significantly less caspase-3 and significantly more NF (NF-L, NF-H and NF-M) in the treatment group than those in the control group at Day 3, 7, and 14 (P<0.05). There was positive correlation between modified Rivlin loxotic plate degree and NF expression, BBB score and NF expression; and negative correlations between BBB score and caspase-3 expression, caspase-3 expression and NF expression. CONCLUSION: Tetramethylpyrazine improves spinal cord healing through regulation of caspase-3 and neurofilament protein expression.

[Effect of low-intensity pulsed ultrasound on the enchondral bone formation in posterolateral lumbar fusion in rabbits].

OBJECTIVE: To observe the effect of daily low-intensity pulsed ultrasound (LIPUs) therapy on improving the enchondral bone formation in lumbar fusion in rabbit models, and to explore its possible mechanism. METHODS: Posterolateral noninstrumented bilateral fusions were performed at the L5 approximately L6 levels in 20 New Zealand rabbits. The autograft iliac bone was implanted on the left side, and the hydroxyapatite bioceramic artificial bone on the right. The rabbits were divided into a treatment group and a control group randomly. One week after the surgery, LIUPs was administered for 20 minutes per day for 4 weeks over the fusion site in the treatment group and false treatment was used in the control group. Post-anterior X-ray photographs were taken to determine the conditions of fusion area, and then, rabbits were killed and the fusion tissues were obtained. Chondrocytes were detected by histological and cytological methods. RESULTS: Compared with the control group, the fusion rate of the treatment group was significantly up-regulated (P<0.05). There was plenty bone trabecula in the fusion area in the treatment group, the number of chondrocytes was also higher than that of the control group (P<0.05), and there was no statistical difference in the number of chondrocytes between the iliac and artificial bone tissues after the treatment(P>0.05). CONCLUSION: Low-intensity pulsed ultrasound therapy improves the endochondral bone formation in the lumbar spine fusion in rabbit models.

[Clinical analysis of the treatment of reserving limb for patients with malignant bone tumor].

OBJECTIVE: To investigate the curative effect of the treatment of reserving limb for patients with malignant bone tumor. METHODS: Forty patients with bone neoplasms underwent the operation of reserving limbs. The neoadjuvant chemotherapy was given to all the patients before the operation. After the ablation of the tumor we used the artificial prosthetic replacement,tumor tissue deactivation and reimplant, variant bone and joint transplantation to reserve the limb. RESULTS: All the patients were followed up. The average follow-up time was 34.8 months. Eight patients got complications. And the fineness rate was 72.5% according to functional evaluation standard of Enneking after surgical treatment of bone tumor. CONCLUSION: The operation of reserving limb was an ideal method to cure the malignant bone tumor. The proper operative treatment with neoadjuvant chemo-therapy is helpful to extend the life of the patients and increase the quality of life.

[Different operation methods for thoracolumbar spinal tuberculosis].

OBJECTIVE: To compare the advantages and disadvantages among the conventional operative methods with the primary anterior debridement, intercentrum auto grafting, and anterior internal fixation method for the thoracolumbar spinal tuberculosis, and to discuss the applying principle. METHODS: Twenty-seven patients with thoracolumbar spinal tuberculosis were treated by the primary anterior debridement, intercentrum auto grafting, and anterior internal fixation, 8 patients were treated by focus debridement and intercentrum auto grafting operations, and the other 22 patients were treated by focus debridement from Feburary 1998 to July 2003, and we analysed the status of the grafting bone fused, the neural function resumed, the malformation rectified and the whole cure ratio. RESULTS: All patients were cured after the follow-up of 15.2 months averagely. Three thoracic spinal and 1 lumbar spinal tuberculosis patients who suffered local infection were cured by debridement and other managements. The time of beginning active movement postoperatively was shortened, and the degrees of kyphosis correction had increased in the anterior internal fixation group compared with the other two groups (P <0.05). But the cost and the bleeding resulted from operation had increased in the anterior internal fixation group compared with the other two groups (P <0.05). CONCLUSION: The conventional and the modern operation methods of thoracolumbar spinal tuberculosis have respective advantages and disadvantages, so that they can complement rather than replace each other.

Hydroxyapatite/tricalcium phosphate matrix scaffold as cell carriers in vitro.

OBJECTIVE: To explore the possibility of hydroxyapatite/tricalcium phosphate (HA/TCP) matrix scaffold as cell carriers in tissue engineering, and to provide a direct evidence for the resurfacing of matrix scaffold. METHODS: Human bone marrow stromal cells (hBMSCs) and umbilical vein endothelial cells (UVECs) were co-cultured on the surface of matrix scaffold. The morphologic characters during culture on HA/TCP matrix scaffold's surfaces were checked by light microscope, fluorescence microscope, and scanning electronic microscope. RESULTS: Both human bone marrow stromal cells and human umbilical vein endothelial cells showed good biocompatibility with HA/TCP matrix scaffold. The cell growth into the micropore of HA/TCP matrix scaffold could be seen under both fluorescence microscopy and scaning electron microscopy. CONCLUSION: HA/TCP matrix scaffold can be used as a vehicle for the cell transplantation.