Expression pattern of MMP-12 in long-term optic nerve regeneration induced by lens injury / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the expression changes of MMP-12 during the long-term axon regeneration induced by the lens injury after the optic nerve clamp trauma in sprague-dawley(SD)rats.METHODS: The optic nerve injury model and lens injury model of SD rats were established, and the 24 experimental animals were divided into control group; lens injury group; optic nerve injury group; lens injury combined with optic nerve injury group, with 6 rats in each group. Reference transcriptome sequencing was used to analyze the expression changes of differentially expressed genes in the injured optic nerve region, and relevant differentially expressed genes with high expression were screened. Quantitative real-time polymerase chain reaction(qRT-PCR)and enzyme-linked immunosorbent assay(ELISA)were used to quantify the expression changes of matrix metalloproteinase-12(MMP-12)in the injured optic nerve region.RESULTS: The Principal Component Analysis of transcriptome sequencing indicated that lens injury combined with optic nerve injury was the principal component of gene expression change. Analysis of gene expression differences showed that the expression of MMP-12 gene was up-regulated in the lens injury combined with optic nerve injury group. The mRNA expression level of MMP-12 in the lens injury combined optic nerve injury group was up-regulated compared with the control group, the optic nerve injury group and the lens injury group at 14d and 21d after successful modeling(P<0.05). At 7, 28d, there was no difference in expression among all groups. The protein expression level of MMP-12 in the lens injury combined with optic nerve injury group was up-regulated compared with the control group and optic nerve injury group at 7, 14 and 21d after successful modeling(P<0.05), and it was up-regulated in the lens injury group combined with optic nerve injury group compared with optic nerve injury group at 21d(P<0.05). At 28d, there was no difference in expression among all groups.CONCLUSION: The up-regulated expression of MMP-12 may be involved in the long-term regeneration of the optic nerve after lens injury.

E.faecium QH06 alleviates TNBS-induced colonic mucosal injury in rats / 南方医科大学学报

OBJECTIVE@#To investigate the effect of Enterococcus faecium QH06 on TNBS-induced ulcerative colitis (UC) in rats and explore the mechanisms in light of intestinal flora and intestinal immunity.@*METHODS@#Thirty-six male Wistar rats were randomized equally into control group, UC model group, and E.faecium QH06 intervention group. The rats in the latter two groups were subjected to colonic enema with 5% TNBS/ethanol to induce UC, followed by treatment with intragastric administration of distilled water or E.faecium QH06 at the dose of 0.21 g/kg. After 14 days of treatment, the rats were examined for colon pathologies with HE staining. The mRNA and protein expression levels of IL-4, IL-10, IL-12, and IFN-γ in the colon tissues were detected using RT-qPCR and ELISA, and the expression of TLR2 protein was detected with immunohistochemistry and ELISA. Illumina Miseq platform was used for sequencing analysis of the intestinal flora of the rats with bioinformatics analysis. The correlations of the parameters of the intestinal flora with the expression levels of TLR2 and cytokines were analyzed.@*RESULTS@#The rats with TNBS- induced UC showed obvious weight loss (P < 0.01) and severe colon tissue injury with high pathological scores (P < 0.01). The protein expression levels of IFN-γ, IL-12, and TLR2 (P < 0.01) and the mRNA expression levels of IFN-γ, IL-12 and IL-10 (P < 0.05) were significantly increased in the colon tissues of the rats with UC. Illumina Miseq sequence analysis showed that in UC rats, the Shannon index (P < 0.05) ACE (P < 0.01)and Chao (P < 0.05) index for the diversity of intestinal flora both decreased with a significantly increased abundance of Enterobacteriaceae (P < 0.01) and a lowered abundance of Burkholderiaceae (P < 0.05). Compared with the UC rats, the rats treated with E. faecium QH06 showed obvious body weight gain (P < 0.05), lessened colon injuries, lowered pathological score of the colon tissue (P < 0.05), decreased protein expressions of IFN- γ, IL- 12, and TLR2 and mRNA expressions of IFN- γ and IL-12 (P < 0.01 or 0.05), and increased protein expressions of IL- 4 (P < 0.05). The Shannon index ACE (P < 0.05) and Chao (P < 0.05) index of intestinal microflora were significantly increased, the abundance of Enterobacteriaceae was lowered and that of Burkholderiaceae and Rikenellaceae was increased in E.faecium QH06- treated rats (P < 0.01 or 0.05). Correlation analysis showed that IFN-γ was positively correlated with the abundance of Enterobacteriaceae, and IFN-γ was negatively correlated with the abundance of Prevotellaceae, Desulfovibrionaceae, norank_o_Mollicutes_RF39 and Clostridiales_vadinBB60_group; TLR2 was negatively correlated with Clostridiales_vadinBB60_group, norank_o_Mollicutes_RF39 and Prevotellaceae.@*CONCLUSION@#E.faecium QH06 can alleviate TNBS-induced colonic mucosal injury in rats, and its effect is mediated possibly by increasing the abundance of SCFA-producing bacteria such as Prevotellaceae and inhibiting abnormal immune responses mediated by TLR2.

Research on the lncRNA MALAT1 promoting the proliferation, migration and angiogenesis of retinal vascular endothelial cells in diabetic retinopathy through the molecular axis of miR-124-3p/SOX7 / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the effect of lncRNA MALAT1 on the proliferation, migration and angiogenesis of retinal vascular endothelial cells and its molecular mechanism.METHODS: The expression levels of lncRNA MALAT1 in plasma of normal control group, diabetic without retinopathy group and diabetic retinopathy group were detected by qPCR and the effect of glucose culture on the expression levels of lncRNA MALAT1 were detected by qPCR too. The expression level of miR-124-3p was detected by qRT-PCR; Western blotting was used to detect the expression level of SOX7; The targeting relationship between lncRNA MALAT1 and miR-124-3p, miR-124-3p and SOX7 were detected by the dual-luciferase reporter system; CCK-8 assay was used to detect cell proliferation activity; Transwell assay was used to detect the migration ability of cells; Angiogenesis of hRMECs cells was measured by in vitro tube formation assay.RESULTS:The expression level of lncRNA MALAT1 in plasma of diabetic retinopathy patients was significantly higher than that of diabetic without retinopathy group and normal control group(P&#x003C;0.001). In vitro glucose culture significantly promoted the expression of lncRNA MALAT1 in hRMECs cells, as well as the proliferation, migration and angiogenesis of hRMECs cells(all P&#x003C;0.05). Knockdown of lncRNA MALAT1 significantly inhibited the proliferation, migration and tubule formation of hRMECs cells(all P&#x003C;0.05). Dual-luciferase reporter gene assay showed that lncRNA MALAT1 targeted with miR-124-3p, and miR-124-3p targeted with SOX7. Overexpression of miR-124-3p significantly inhibited the proliferation, migration and tubule formation of hRMECs cells(all P&#x003C;0.05). Overexpression of lncRNA MALAT1+miR-124-3p, miR-124-3p+SOX7, and knockdown of lncRNA MALAT1+overexpression of SOX7 all significantly eliminated the inhibitory effect of hRMECs cells(all P&#x003C;0.05).CONCLUSION: lncRNA MALAT1 promote the proliferation, migration and angiogenesis of retinal endothelial cells in diabetic retinopathy by down-regulating the negative regulation of miR-124-3p on SOX7. Therefore, abnormal upregulation of lncRNA MALAT1 in patients with diabetic retinopathy is a potential biomarker.

Effect of salidroside on the expression of connexin43 in the corpus cavernosum smooth muscle cells of hypoxic rats / 中华男科学杂志

<p><b>Objective</b>To investigate the effect of salidroside on the expression of the connexin43 (Cx43) protein in the corpus cavernosum smooth muscle cells (CCSMCs) of hypoxic SD rats.</p><p><b>METHODS</b>CCSMCs were cultured in vitro and identified by immunofluorescence staining. The cells were divided into six groups: normal control (21% O2), hypoxia (1% O2), hypoxia+salidroside (HS) 8 μg/ml,HS 16 μg/ml, HS 32 μg/ml, and HS 64 μg/ml, and cultured for 48 hours. Then the relative expression of Cx43 in different groups was detected by Western blot.</p><p><b>RESULTS</b>The in vitro cultured CCSMCs grew well and 90% of the cells showed positivity for α-SMA and desmin on immunohistochemistry. Salidroside ≤64 μg/ml produced no obvious toxicity on the CCSMCs. The expressions of Cx43 and phosphorylated proteins were dramatically increased in the hypoxia group as compared with the normal control (P<0.01 and P<0.05). The HS groups all showed significantly higher expression of Cx43 than the hypoxia group (P<0.01), but the phosphorylation rate of the Cx43 proteins was remarkably decreased (P<0.01).</p><p><b>CONCLUSIONS</b>Hypoxia increases the expression of Cx43 in the CCSMCs of SD rats. Salidroside ≤64 μg/ml cannot reverse the hypoxia-induced change but can reduce the dephosphorylation of Cx43 in CCSMCs. It is deduced that salidroside can protect CCSMCs by decreasing the phosphorylation of Cx43 and suppressing hypoxia-induced formation of the gap junction channel.</p>

Inhibitory effect of salidroside on hypoxia-induced apoptosis of corpus cavernosum smooth muscle cells in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effect of salidroside on hypoxia-induced apoptosis of corpus cavernosum smooth muscle cells (CCSMCs) in rats.</p><p><b>METHODS</b>Rat CCSMCs were cultured in vitro by the enzyme digestion method and identified by immunofluorescent staining of anti-alpha-SMA and anti-Desmin. The non-toxic dose of salidroside was determined by MTT assay. Low-oxygen mixed gas (1% O2, 5% CO2, and 94% N2) was piped into a modular incubator chamber to induce hypoxia. The CCSMCs were divided into a normal, a hypoxia, and a 32 microg/mL salidroside intervention group. The apoptosis of the CCSMCs was detected by flow cytometry and the expression of the caspase-3 protein determined by Western blot.</p><p><b>RESULTS</b>The majority of the CCSMCs were positive for alpha-SMA and Desmin at immunofluorescent staining. Salidroside at < 32 microg/ml produced no obvious toxicity to CCSMCs. Compared with the normal control group, the rates of early and late apoptosis of CCSMCs were both increased significantly in the hypoxia group ([12.77 +/-1.41]% vs [18.69 +/- 1.29]%, P < 0.01 and [14.63 +/- 2.00]% vs [21.03 +/- 1.530]% , P < 0.05). Western blot showed a markedly increased expression of cleaved caspase-3 (P < 0.01). Intervention with 32 microg/ml salidroside significantly reduced hypoxia-induced early apoptosis of CCSMCs ([13.46% +/- 1.87]%, P < 0.01) and decreased the expression of cleaved caspase-3 (P < 0.01).</p><p><b>CONCLUSION</b>Salidroside can reduce the expression of cleaved caspase-3 and inhibit hypoxia-induced apoptosis of CCSMCs in rats.</p>

Chemical constituents from safflower injection and their bioactivity / 中国中药杂志

The chemical constituents of Safflower injection were isolated and purified by polyamide, silica gel, Sephadex LH-20, ODS column chromatographies and preparative HPLC. As a result, sixteen compounds have been isolated. Based on the spectral data analysis, their structures were elucidated as scutellarin (1), kaempferol-3-O-β-rutinoside(2), hydroxysafflor yellow A(3), rutin (4), coumalic acid(5), adenosine(6), syringoside(7), (3E)-4-(4'-hydroxyphenyl)-3-buten-2-one(8), (8Z)-decaene-4, 6-diyne-1-Oβ-D-glucopyranoside(9), 4-hydroxybenzaldehyde (10), (2E, 8E) -tetradecadiene-4, 6-diyne-1, 12, 14-triol-1-O-β-D-glucopyranoside (11), kaem-pferol-3-O-β-sophorose (12), uridine (13), roseoside (14), cinnamic acid (15), and kaempferol (16). Compounds 1,2,7,9,11 and 12 were isolated from the Safflower injection for the first time. The anti-platelet aggregation activities of the isolated compounds were assayed. The results indicated all tested compounds exhibited potent activity except for 5, while 2, 3, 9 and 12 showed strong activity against platelet aggregation.

Single-segment pedicle screw fixation for the treatment of thoracolumbar fractures through the gap of paravertebral muscles / 中国骨伤

<p><b>OBJECTIVE</b>To compare the clinical effect of thoracolumbar fractures between single-segment pedicle screw fixation approach for the gap of paravertebral muscles and double-segment pedicle screw fixation approach for the stripping of paravertebral muscles.</p><p><b>METHODS</b>From September 2008 to January 2010, 65 patients with incomplete compressed thoracolumbar fractures or burst thoracolumbar fractures with unilateral endplate injury were randomly divided into two groups. Thirty patients were treated with single-segment pedicle screw fixation through the gap of paravertebral muscles (treatment group). Thirty-five patients were treated with double-segment pedicle screw fixation through the stripping of the paravertebral muscles (control group). All the internal fixations were taken out during 10-12 months after operation. Operative time, perioperative blood loss volume and postoperative drainage volume were compared between two groups. At final follow-up, the change of neurological ASIA grade were recorded; and postoperative 5 days and final follow-up, compared Denis classification of lumbar and back pain between two groups; and analyzed the sagittal index and compressibility of anterior border of vertebral body by X -ray lateral projection.</p><p><b>RESULTS</b>All patients were follow-up from 14 to 22 months with an average of 18.3 months. No postoperative infection, secondary spinal cord injury was found. One case of control group occurred internal fixation breakage at the 11th month after operation and other internal fixation no loosening. There was no significant difference in operative time, the recovery of neurological function between the two groups (P > 0.05). Perioperative blood loss volume and postoperative drainage volume of treatment group was less than that of control group (P < 0.01). And in Denis classification of lumbar and back pain, the treatment group recovered more quickly, and the residual pain of lumbar and back was less than that of control group (P < 0.01). Postoperative posterior salient and compression of anterior border of vertebral body improved in two groups (P < 0.01), there was no significant difference in degree of improvement between two groups (P > 0.05); but both loss existed at final follow-up (P < 0.01), there was no significant difference in loss of posterior salient between two groups (P > 0.05). In the treatment group, the loss of rectify of anterior border of vertebral body existed, but it was less than that of the control group.</p><p><b>CONCLUSION</b>In the premise of strict controlling surgery indications, the treatment of thoracolumbar fractures with single-segment pedicle screw fixation through the gap of paraspinal muscles, can effectively recover the height of vertebral body and rectify posterior salient, and reduce the fixed segment. Compared with the traditional operative method of double-segment pedicle screw fixation through the stripping of paraspinal muscle, it can obviously reduce the operation wound and the bleeding, lessen the pain of lumbar and back. And the recent clinical effect is satisfied.</p>