Identification and validation of iron metabolism genes in osteoporosis.

BACKGROUND: Osteoporosis is the most common metabolic bone disease in humans. Exploring the expression difference of iron metabolism-related genes in osteoporosis can provide a new target for diagnosis and treatment. METHODS: First, we used online databases to identify differentially expressed genes (DEGs) related to iron metabolism in patients with osteoporosis. The differential genes were comprehensively analyzed by bioinformatics method (GO, KEGG, GSEA, immune infiltration analysis, PPI). The expression levels of hub genes and important signaling pathways were verified by qRT-PCR and Western blotting. RESULTS: A total of 23 iron metabolism-related genes with significant differences were identified, which were enriched in "regulation of protein dephosphorylation" and "negative regulation of protein dephosphorylation". The GSEA results, heme metabolism and Myc targets v1 were among the top two pathways, both upregulated. The immune infiltration analysis revealed that the expressions of genes such as ABCA5, D2HGDH, GNAI2, and CTSW were correlated with the infiltration degree of significantly different cells. The PPI network contained 12 differentially expressed iron metabolism-related genes. Additionally, YWHAE, TGFB1, PPP1R15A, TOP2A, and CALR were mined as hub genes using the Cytoscape software. qRT PCR showed that the expression of TGF-ß1, YWHAE, TOP2A and CALR increased. We also verified the expression of related proteins and genes in the oxidative stress signaling pathway by qRT PCR and Western blotting. The results showed that Mob1, YAP and TAZ molecules were highly expressed at the gene and protein levels. CONCLUSIONS: These differentially expressed iron metabolism-related genes could provide new potential targets for the diagnosis and treatment of osteoporosis.

MicroRNA-582-3p regulates osteoporosis through regulating homeobox A10 and osteoblast differentiation.

OBJECTIVE: Recent studies have demonstrated that micro RNAs (miRNAs) are involved in bone formation and bone cell differentiation, but the role of miR-582-3p in osteoporosis is unclear. We want to study the mechanism of miR-582-3p on osteogenic differentiation. METHOD: The expression of miR-582-3p and homeobox (Hox) A10 were analyzed by quantitative RT-PCR. The expression levels of HOXA10 protein were determined by Western blot. The target of HOXA10 was identified by bioinformatics and luciferase reporter gene assay. RESULTS: The results showed that miR-582-3p was up-regulated in OP tissues and down-regulated in osteogenic differentiated C2C12 cells compared with that in the control group. Overexpression of miR-582-3p resulted in reduced expression levels of osteocalcin (OC), alkaline phosphatase (ALP), and collagen, type I, α1 (COL1A1). miR-582-3p had a potential binding site with HOXA10. Moreover, miR-582-3p inhibited the expression of HOXA10, and overexpression of HOXA10 reduced the effect of miR-582-3p on osteoblast markers. HOXA10 was the target gene of miR-582-3p, which could inhibit the expression of HOXA10. Furthermore, HOXA10 reduced the role of miR-582-3p in osteoblast markers. miR-582-3p inhibited the development of osteoporosis by regulating HOXA10 and osteoblast differentiation. CONCLUSIONS: miR-582-3p may be a therapeutic target of osteoporosis treatment.

The efficacy of bloodletting therapy in patients with acute gouty arthritis: A systematic review and meta-analysis.

BACKGROUND: Bloodletting therapy (BLT) is widely used to relieve acute gouty arthritis (AGA). However, limited evidence-based reports exist on the effectiveness and safety of BLT. This systematic review aims to evaluate the feasibility and safety of BLT in treating AGA. METHODS: Seven databases were exhaustively screened from the date of establishment to July 31, 2020, irrespective of the publication source and language. The included articles were evaluated for bias risk by using the Cochrane risk of bias assessment tool. All statistical analyses were done with Review Manager 5.3. RESULTS: Twelve studies involving 894 participants were included for the final analysis. Our meta-analysis revealed that BLT was highly effective in relieving pain (MD = -1.13, 95% CI [-1.60, -0.66], P < 0.00001), with marked alterations in the total effective (RR = 1.09, 95% [1.05, 1.14], P < 0.0001) and curative rates (RR = 1.37, 95%CI [1.17, 1.59], P < 0.0001). In addition, BLT could dramatically reduce serum C-reactive protein (CRP) level (MD = -3.64, 95%CI [-6.72, -0.55], P = 0.02). Both BLT and Western medicine (WM) produced comparable decreases in uric acid (MD = -18.72, 95%CI [-38.24, 0.81], P = 0.06) and erythrocyte sedimentation rate (ESR) levels (MD = -3.01, 95%CI [-6.89, 0.86], P = 0.13). Lastly, we demonstrated that BLT was safer than WM in treating AGA (RR = 0.36, 95%CI [0.13, 0.97], P = 0.04). CONCLUSION: BLT is effective in alleviating pain and decreasing CRP level in AGA patients with a lower risk of evoking adverse reactions.

Effect of intensity-modulated radiation therapy on sciatic nerve injury caused by echinococcosis.

Conventional radiotherapy has a good killing effect on femoral echinococcosis. However, the sciatic nerve around the lesion is irreversibly damaged owing to bystander effects. Although intensity-modulated radiation therapy shows great advantages for precise dose distribution into lesions, it is unknown whether intensity-modulated radiation therapy can perfectly protect the surrounding sciatic nerve on the basis of good killing of femoral echinococcosis foci. Therefore, this study comparatively analyzed differences between intensity-modulated radiation therapy and conventional radiotherapy on the basis of safety to peripheral nerves. Pure-breed Meriones meridiani with bilateral femoral echinococcosis were selected as the research object. Intensity-modulated radiation therapy was used to treat left femoral echinococcosis of Meriones meridianus, while conventional radiotherapy was used to treat right femoral echinococcosis of the same Meriones meridianus. The total radiation dose was 40 Gy. To understand whether intensity-modulated radiation therapy and conventional radiotherapy can kill femoral echinococcosis, trypan blue staining was used to detect pathological changes of bone Echinococcus granulosus and protoscolex death after radiotherapy. Additionally, enzyme histochemical staining was utilized to measure acid phosphatase activity in the protoscolex after radiotherapy. One week after radiotherapy, the overall structure of echinococcosis in bilateral femurs of Meriones meridiani treated by intensity-modulated radiation therapy disappeared. There was no significant difference in the mortality rate of protoscoleces of Echinococcus granulosus between the bilateral femurs of Meriones meridiani. Moreover, there was no significant difference in acid phosphatase activity in the protoscolex of Echinococcus granulosus between bilateral femurs. To understand the injury of sciatic nerve surrounding the foci of femoral echinococcosis caused by intensity-modulated radiation therapy and conventional radiotherapy, the ultrastructure of sciatic nerves after radiotherapy was observed by transmission electron microscopy. Additionally, apoptosis of neurons was examined using a terminal-deoxynucleotidyl transferase-mediated dUTP nick end labeling assay, and expression of Bcl-2 and Bax in sciatic nerve tissue was detected by immunohistochemical staining and western blot assay. Our results showed that most neurons in the left sciatic nerve of Meriones meridiani with echinococcosis treated by intensity-modulated radiation therapy had reversible injury, and there was no obvious apoptosis. Compared with conventional radiotherapy, the number of apoptotic cells and Bax expression in sciatic nerve treated by intensity-modulated radiation therapy were significantly decreased, while Bcl-2 expression was significantly increased. Our findings suggest that intensity-modulated radiation therapy has the same therapeutic effect on echinococcosis as conventional radiotherapy, and can reduce apoptosis of the sciatic nerve around foci caused by radiotherapy. Experiments were approved by the Animal Ethics Committee of People's Hospital of Xinjiang Uygur Autonomous Region, China (Approval No. 20130301A41) on March 1, 2013.

Contralateral Electroacupuncture Relieves Chronic Neuropathic Pain in Rats with Spared Nerve Injury.

BACKGROUND Acupuncture and electroacupuncture (EA) are widely applied in the treatment of various conditions, including pain. Acupuncture stimulation is applied not only in areas close to pain sites, but also in distal regions or on the contralateral side of the body. Identifying which acupuncture paradigms produce best therapeutic effects is of clinical significance. MATERIAL AND METHODS Spared nerve injury (SNI) was applied to establish a rat model of neuropathic pain. We applied 14 sessions of EA (BL 60 and BL 40, 1-2 mA, and 2 Hz, 30 min per session) every other day from days 3 to 29 after surgery on the contralateral or ipsilateral side of pain. von Frey hair was applied to examine mechanical allodynia in the SNI model and analgesic effects of EA. All experimental procedures were approved by the Animal Care and Use Committee of our university, according to the guidelines of the International Association for the Study of Pain. RESULTS SNI produced significant and long-lasting mechanical allodynia (p<0.001) in injured paws. Repeated EA on the contralateral side of the pain significantly attenuated mechanical allodynia from 14 days after surgery (p<0.05). By contrast, ipsilateral EA did not show analgesic effects (p>0.05). CONCLUSIONS These findings indicate that contralateral EA is superior to local EA in some types of pain disorders. Further investigations are needed for a more comprehensive understanding of the central mechanisms of acupuncture.

MicroRNA-194-3p inhibits the metastatic biological behaviors of spinal osteosarcoma cells by the repression of matrix metallopeptidase 9.

Osteosarcoma is the most common primary malignant bone tumor, but only 3%-5% of cases occur in the spine. Spinal osteosarcoma presents a significant challenge, and most patients die in spite of aggressive surgery. MicroRNAs (miRNAs) are small noncoding RNAs that have a pivotal role in the post-transcriptional regulation of gene expression. The aim of this study was to investigate the role of miR-194-3p and to identify its potential mechanism in spinal osteosarcoma. Here, spinal osteosarcoma tissues showed down-regulated expression of miR-194-3p compared to adjacent non-tumorous tissues. The level of miR-194-3p was negatively correlated with metastasis in patients with spinal osteosarcoma. MiR-194-3p over-expression in spinal osteosarcoma cells significantly inhibited cell migration and invasion in vitro. Furthermore, mechanistic analyses showed that MMP-9 (matrix metallopeptidase 9) is a direct target of miR-194-3p, and the ectopic expression of miR-194-3p inhibits MMP-9 expression by directly binding to the 3'-untranslated region (3'-UTR) of the MMP-9 gene. In summary, our results demonstrate that miR-194-3p suppresses migration and invasion of spinal osteosarcoma cells by targeting MMP-9, indicating miR-194-3p may serve as a promising novel target for spinal osteosarcoma therapy.

Regulating the Balance of Th17/Treg via Electroacupuncture and Moxibustion: An Ulcerative Colitis Mice Model Based Study.

AIM: To investigate the relationship between the effects of electroacupuncture/moxibustion and the balance of Th17/Treg in treating ulcerative colitis (UC) and to preliminary compare the effects of the above two methods. METHODS: DSS-induced UC mice were treated by electroacupuncture and moxibustion. Disease activity index (DAI) was scored; intestinal pathological structure and ultrastructure were observed. The levels of IL-2, IL-6, IL-10, IL-17A, IL-17F, and TGF-ß in plasma were measured by ELISA. The percentages of Treg and Th17 in spleen lymphocytes were analyzed by flow cytometry. Also, the expressions of TLR2, TLR4, RORγt, and FOXP3 in the distal colon were detected by immunohistochemistry or western blot. RESULTS: Both electroacupuncture and moxibustion can relieve UC. These effects are further supported by ELISA results. In addition, the ratio of Treg and Th17 in spleen lymphocytes and the expression of TLR2 and TLR4 are significantly improved. Also, the expression of RORγt and FOXP3 in distal colon were improved. Besides, the effect of moxibustion is better than that of electroacupuncture on TLR2, TLR4, and FOXP3 expression (P < 0.05). CONCLUSION: Both electroacupuncture and moxibustion may ameliorate UC by regulating the balance of Th17/Treg. Whether moxibustion has better efficacy than electroacupuncture needs further study.

Combined use of Y-tube conduits with human umbilical cord stem cells for repairing nerve bifurcation defects.

Given the anatomic complexity at the bifurcation point of a nerve trunk, enforced suturing between stumps can lead to misdirection of nerve axons, thereby resulting in adverse consequences. We assumed that Y-tube conduits injected with human umbilical cord stem cells could be an effective method to solve such problems, but studies focused on the best type of Y-tube conduit remain controversial. Therefore, the present study evaluated the applicability and efficacy of various types of Y-tube conduits containing human umbilical cord stem cells for treating rat femoral nerve defects on their bifurcation points. At 12 weeks after the bridging surgery that included treatment with different types of Y-tube conduits, there were no differences in quadriceps femoris muscle weight or femoral nerve ultrastructure. However, the Y-tube conduit group with longer branches and a short trunk resulted in a better outcome according to retrograde labeling and electrophysiological analysis. It can be concluded from the study that repairing a mixed nerve defect at its bifurcation point with Y-tube conduits, in particular those with long branches and a short trunk, is effective and results in good outcomes.

The downregulation of miR­125a­5p functions as a tumor suppressor by directly targeting MMP­11 in osteosarcoma.

Osteosarcoma is one of the most common primary malignant bone cancers in juveniles and adults. Increasingly, reports indicate that microRNAs (miRNAs) may provide novel therapeutic targets for cancer treatment. The aim of the present study was to investigate the expression of miR­125a­5p and to identify its functional significance in osteosarcoma. This indicated that miR­125a­5p was downregulated in osteosarcoma tissue and cell lines using reverse transcription­quantitative polymerase chain reaction. Following transfection with miR­125a­5p mimics or the negative control, cell migration, invasion and epithelial­mesenchymal transition (EMT) assays were conducted in osteosarcoma cells. These results indicated that the overexpression of miR­125a­5p resulted in inhibited osteosarcoma cell migration, invasion and EMT in vitro. Furthermore, mechanistic studies showed that matrix metallopeptidase­11 (MMP­11), was a direct target of miR­125a­5p in osteosarcoma. Taken together, the data demonstrate that miR­125a­5p functions as a tumor suppressor gene and serves an important role in inhibiting osteosarcoma cell migration, invasion and EMT by targeting MMP­11.

Randomised controlled trial of contralateral manual acupuncture for the relief of chronic shoulder pain.

OBJECTIVE: To explore the effects of contralateral manual acupuncture (MA) on patients with chronic shoulder pain. METHODS: Eighty patients with chronic shoulder pain were randomly allocated to receive contralateral MA (n=38) for 4 weeks or to remain on a waiting list while receiving conventional orthopaedic therapy (n=42). Visual analogue scale (VAS) scores were taken as the primary outcome measure and used for a priori power calculation. Secondary outcome measures for the assessment of shoulder mobility and quality of life included the Jobe test, the Constant-Murley (CM) score, the Disabilities of the Arm, Shoulder and Hand (DASH) score, and the 36 item Short Form Health Survey (SF-36). RESULTS: Intention-to-treat (ITT) analysis demonstrated significant pain relief with contralateral acupuncture, with mean differences in VAS scores compared to the waiting list group of -19.4 (-28.0 to -10.8) at 2 weeks, -40.4 (-49.0 to -31.8) at 4 weeks, -41.1 (-49.7 to -32.5) at 8 weeks, and -40.9 (-49.5 to -32.3) at 16 weeks. CM and DASH scores were also improved at all time points (p<0.01). Shoulder mobility, physical functioning, social functioning and mental health components of the SF-36 were also improved by contralateral acupuncture at 8 weeks. No significant adverse effects were observed. CONCLUSIONS: These results demonstrate beneficial effects of contralateral acupuncture in the treatment of chronic shoulder pain, both in terms of pain and function. Future research is required to compare directly the effects of local and contralateral acupuncture and to quantify the specific and non-specific effects. TRIAL REGISTRATION NUMBER: NCT01733914.

Phylogenetic studies of Hylopetes alboniger based on complete mitochondrial DNA sequences.

The complete mitochondrial genome sequence of the Hylopetes alboniger was sequenced and reported for the first time using muscle tissue. The mitochondrial genome is 16,584 bp in size, including 22 tRNA genes, 13 protein-coding genes, 12S rRNA, 16S rRNA, and one control region. Phylogenetic analysis was performed using whole mitogenome sequences with other 15 closely related taxa to assess their phylogenetic relationship. This mitochondrial genome sequence will provide a better understanding for H. alboniger evolution in the future.

Three-dimensional conformal intensity-modulated radiation therapy of left femur foci does not damage the sciatic nerve.

During radiotherapy to kill femoral hydatid tapeworms, the sciatic nerve surrounding the focus can be easily damaged by the treatment. Thus, it is very important to evaluate the effects of radiotherapy on the surrounding nervous tissue. In the present study, we used three-dimensional, conformal, intensity-modulated radiation therapy to treat bilateral femoral hydatid disease in Meriones meridiani. The focus of the hydatid disease on the left femur was subjected to radiotherapy (40 Gy) for 14 days, and the right femur received sham irradiation. Hematoxylin-eosin staining, electron microscopy, and terminal deoxynucleotidyl transferase-dUTP nick end labeling assays on the left femurs showed that the left sciatic nerve cell structure was normal, with no obvious apoptosis after radiation. Trypan blue staining demonstrated that the overall protoscolex structure in bone parasitized with Echinococcus granulosus disappeared in the left femur of the animals after treatment. The mortality of the protoscolex was higher in the left side than in the right side. The succinate dehydrogenase activity in the protoscolex in bone parasitized with Echinococcus granulosus was lower in the left femur than in the right femur. These results suggest that three-dimensional conformal intensity-modulated radiation therapy achieves good therapeutic effects on the secondary bone in hydatid disease in Meriones meridiani without damaging the morphology or function of the sciatic nerve.

[Assessment of percutaneous vertebroplasty and percutaneous kyphoplasty for treatment of senile osteoporotic vertebral compression fractures].

OBJECTIVE: To investigate the clinical effects and safety of vertebroplasty and kyphoplasty for the senile osteoporotic vertebral compression fractures. METHODS: From December 2004 to June 2008, 28 patients (40 vertebrae) with osteoporotic vertebral compression fractures were treated with percutaneous vertebroplasty (PVP group), there were 11 males (14 vertebrae) and 17 females (26 vertebrae), with an average age of 72 years (ranged, 70 to 91 years). The fracture site of vertebral body was from T5 to L5. Other 31 patients (43 vertebrae) were treated with percutaneous kyphoplasty (PKP group), there were 13 males (18 vertebrae) and 18 females (25 vertebrae), with an average age of 74 years (ranged, 70 to 92 years). The fracture site of vertebral body was from T4 to L5. Theapeutic effects of all the patients were observed after operation, the item included visual analog scale (VAS), Oswestry score, Cobb angle, the height of anterior and middle column vertebra, and the incidence of cement leakage. RESULTS: There was significant decrease in VAS score and Oswestry score after operation in both groups, there was significant difference between preoperative and postoperative (P < 0.01). There was no significant difference in VAS score and Oswestry score between two groups (P > 0.05). There was no significant difference in Cobb angles and the height of anterior and middle column vertebra between two groups before and after operation (P > 0.05). Among them, there were 3 vertebrae cement leakage in PVP group (7.5%) and 2 in PKP group (4.7%), but no clinical sympton occurred. There was no significant difference in the incidence of cement leakage between two groups (P > 0.05). CONCLUSION: Vertebroplasty and kyphoplasty are safe and effective methods in treating senile osteoporotic vertebral compression fractures, which can restore the height of fractured vertebra, relieve pain and improve function as well as minimize the incidence of cement leakage.

[Systematic review of the earlier safety of closed suction drainage for knee arthroplasty].

OBJECTIVE: To evaluate the earlier safety of closed suction drainage systems for knee arthroplasty. METHODS: All randomised or quasi-randomised trials comparing the use of closed suction drainage systems with no drainage systems for all types of knee arthroplasty were searched. The trials' quality were assessed, and data was extracted. Where appropriate, results of comparable studies were pooled. RESULTS: Twelve studies involving 1122 participants with 1191 surgical wounds were identified. The methodology of the studies identified and included in the analysis varied considerably. Pooling of results indicated no statistically significant difference in the incidence of wound haematoma, dehiscence or deep vein thrombosis between those allocated to drains and the un-drained wounds. Blood transfusion was required more frequently in those who received drains. No difference between the groups was seen with respect to wound infection and the range of movement of the joint after surgery. CONCLUSIONS: There is insufficient evidence from randomised trials to support the routine use of closed suction drainage in knee arthroplasty. It results in an increase in the number of patients requiring blood transfusion. However, because of the poor trial methodology of many of the studies and the inadequate reporting of outcomes, further randomized trials on this topic are justified.