Photobiomodulation Increases M2-Type Polarization of Macrophages by Inhibiting Versican Production After Spinal Cord Injury.

Spinal cord injury (SCI) is a catastrophic accidence with little effective treatment, and inflammation played an important role in that. Previous studies showed photobiomodulation (PBM) could effectively downregulate the process of inflammation with modification of macrophage polarization after SCI; however, the potential mechanism behind that is still unclear. In the presented study, we aimed to investigate the effect of PBM on the expression level of versican, a matrix molecular believed to be associated with inflammation, and tried to find the mechanism on how that could regulate the inflammation process. Using immunofluorescence technique and western blot, we found the expression level of versican is increased after injury and markedly downregulated by irradiation treatment. Using virus intrathecal injection, we found the knock-down of versican could produce the effect similar to that of PBM and might have an effect on inflammation and macrophage polarization after SCI. To further verify the deduction, we peptide the supernatant of astrocytes to induce M0, M1, and M2 macrophages. We found that the versican produced by astrocytes might have a role on the promotion of M2 macrophages to inflammatory polarization. Finally, we investigated the potential pathway in the regulation of M2 polarization with the induction of versican. This study tried to give an interpretation on the mechanism of inflammation inhibition for PBM in the perspective of matrix regulation. Our results might provide light on the inflammation regulation after SCI.

Photobiomodulation provides neuroprotection through regulating mitochondrial fission imbalance in the subacute phase of spinal cord injury.

Increasing evidence indicates that mitochondrial fission imbalance plays an important role in delayed neuronal cell death. Our previous study found that photobiomodulation improved the motor function of rats with spinal cord injury. However, the precise mechanism remains unclear. To investigate the effect of photobiomodulation on mitochondrial fission imbalance after spinal cord injury, in this study, we treated rat models of spinal cord injury with 60-minute photobiomodulation (810 nm, 150 mW) every day for 14 consecutive days. Transmission electron microscopy results confirmed the swollen and fragmented alterations of mitochondrial morphology in neurons in acute (1 day) and subacute (7 and 14 days) phases. Photobiomodulation alleviated mitochondrial fission imbalance in spinal cord tissue in the subacute phase, reduced neuronal cell death, and improved rat posterior limb motor function in a time-dependent manner. These findings suggest that photobiomodulation targets neuronal mitochondria, alleviates mitochondrial fission imbalance-induced neuronal apoptosis, and thereby promotes the motor function recovery of rats with spinal cord injury.

Potential targets and mechanisms of photobiomodulation for spinal cord injury.

As a classic noninvasive physiotherapy, photobiomodulation, also known as low-level laser therapy, is widely used for the treatment of many diseases and has anti-inflammatory and tissue repair effects. Photobiomodulation has been shown to promote spinal cord injury repair. In our previous study, we found that 810 nm low-level laser therapy reduced the M1 polarization of macrophages and promoted motor function recovery. However, the mechanism underlying this inhibitory effect is not clear. In recent years, transcriptome sequencing analysis has played a critical role in elucidating the progression of diseases. Therefore, in this study, we performed M1 polarization on induced mouse bone marrow macrophages and applied low-level laser therapy. Our sequencing results showed the differential gene expression profile of photobiomodulation regulating macrophage polarization. We analyzed these genes using gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses. Networks of protein-protein interactions and competing RNA endogenous networks were constructed. We found that photobiomodulation inhibited STAT3 expression through increasing the expression of miR-330-5p, and that miR-330-5p binding to STAT3 inhibited STAT3 expression. Inducible nitric oxide synthase showed trends in changes similar to the changes in STAT3 expression. Finally, we treated a mouse model of spinal cord injury using photobiomodulation and confirmed that photobiomodulation reduced inducible nitric oxide synthase and STAT3 expression and promoted motor function recovery in spinal cord injury mice. These findings suggest that STAT3 may be a potential target of photobiomodulation, and the miR-330-5p/STAT3 pathway is a possible mechanism by which photobiomodulation has its biological effects.

PKM2 regulates hepatocellular carcinoma cell epithelial-mesenchymal transition and migration upon EGFR activation.

Pyruvate kinase isozyme type M2(PKM2) was first found in hepatocellular carcinoma(HCC), and its expression has been thought to correlate with prognosis. A large number of studies have demonstrated that epithelial-mesenchymal transition (EMT) is a crucial event in hepatocellular carcinoma (HCC) and associated metastasis, resulting in enhanced malignancy of HCC. However, the roles of PKM2 in HCC EMT and metastasis remain largely unknown. The present study aimed to determine the effects of PKM2 in EGF-induced HCC EMT and elucidate the molecular mechanisms in vitro. Our results showed that EGF promoted EMT in HCC cell lines as evidenced by altered morphology, expression of EMT-associated markers, and enhanced invasion capacity. Furthermore, the present study also revealed that nuclear translocation of PKM2, which is regulated by ERK pathway, regulated ß-catenin-TCF/LEF-1 transcriptional activity and associated EMT in HCC cell lines. These discoveries provide evidence of novel roles of PKM2 in the progression of HCC and potential therapeutic target for advanced cases.

[Expression of protein gene product 9.5, vasoactive intestinal peptide and neuropeptide Y in the mucosa of the neovagina after sigmoid colon vaginoplasty].

OBJECTIVE: To investigate re-innervation in the neovaginal mucosa of patients underwent sigmoid colon vaginoplasty in treatment of Mayer-Rokitansky-Kistner-Hauser Syndrome (MRKHS). METHODS: Biopsies in the upper third of the posterior neovagina were taken in 20 patients treated by sigmoid colon vaginoplasty at 1, 2 and 3 years after surgery, respectively. Protein gene product 9.5 (PGP 9.5), vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY) were detected by immunohistochemical method and compared with those in intact sigmoid colon mucosa. RESULTS: (1) Density of nerve fiber: abundant distribution of PGP 9.5 nerve fibers were observed in the mucosal muscle layer, submucosa, and smooth muscle layer of the neovagina. The nerve fibers of VIP and NPY immunoreactivity were mainly distributed around blood vessels and in the smooth muscles. In the neovagina, the density of nerve fibers of PGP 9.5 of 17 ± 6 were much more than VIP of 2.9 ± 1.0 and NPY of 2.5 ± 0.8 significantly (P < 0.05). (2) Expression of PGP 9.5 in neovagina: at 1 year after surgery, PGP 9.5 positive expression of 14 ± 4 was significantly lower in the neovagina than 28 ± 7 in the intact sigmoid colon (P < 0.05). However, after 2 to 3 years, its expression displayed an upgrade tendency in the neovagina and was significantly higher at the 3 year postoperatively than that at the 1 years postoperatively (22 ± 7 vs. 14 ± 4, P < 0.05). The changes were much more obvious in submucosa. (3) The expression of VIP and NPY in neovagina: at 1 year after surgery, VIP and NPY positive nerve fibers were also decreased in the neovagina when compared with those in the intact sigmoid colon (2.3 ± 0.7 vs. 5.3 ± 1.4, P < 0.05; 2.5 ± 1.1 vs. 5.5 ± 1.1, P < 0.05). At 2 to 3 years after surgery, the positive VIP fiber showed initially decreased and subsequently increased tendency. The density of VIP of 3.7 ± 0.7 in the neovagina at 3 years postoperatively was higher than 2.3 ± 0.7 at 1 years postoperatively (P < 0.05). No significant up-regulation was observed in NPY-positive expression in the neovagina within 3 years after operation. CONCLUSIONS: Distribution of sensory PGP 9.5, VIP and NPY immunoreactive nerve fibers was similar to the pattern observed within the intact sigmoid colon wall. The number of nerve fibers in the neovagina decreased after surgery and then increased subsequently within 3 years after surgery.

[Expression of occludin and zona occluden-1 and their morphologic changes in vagina mucosal cells in patients undergoing vaginal construction by using sigmoid colon].

OBJECTIVE: In order to evaluate the changes of neovaginal defense function in the sight of physical barrier by investigating the changes in the histological morphology and expression of occludin, zona occluden-1 (ZO-1) in mucosal cells of the neovagina in patients undergoing vaginal construction by sigmoid colon. METHODS: Fifteen cases with neovagina constructed by sigmoid colon matched with 15 cases' vaginal mocusa as control specimens were observed with light microscopic and electron microscopic methods. At the same time, the expression of occludin and ZO-1 were semiquantitatively measured by streptavidin-perosidase (SP) immunohistochemical method. RESULTS: (1) Squamous metaplasia was observed in colonic mucosa from lower one-third of the artificial vagina in 10 cases (67%, 10/15). (2) The intercellular tight junctions of neovaginal mucosa became shorter and wider, the number of them were decreased or disappeared by electron microscopic study. (3) In upper neovagina, the positive rates of occludin and ZO-1 expression in mucosal cells were 80% (12/15) and 73% (11/15), respectively. In lower neovaginal mucosa where squamous metaplasia occurred, the positive rates of two protein expression in cells were both 100% (10/10) and also was 100% (15/15) in control group, which showed significant difference (P < 0.05, P < 0.01). In lower neovaginal mucosa where squamous metaplasia hadn't occurred, the positive expression of two protein in cells (4/5, 2/5) were significantly lower than those in lower neovaginal mucosa where squamous metaplasia had happened 100% (10/10), 100% (10/10) and in control group [100% (15/15), 100% (15/15)] (P < 0.05), while there were no significantly difference in positive expression of two protein in cells between this group and upper neovagina (P > 0.05). CONCLUSIONS: The squamous metaplasia had occurred in lower neovaginal mucosa. The decreased expression of occludin and ZO-1 in upper and lower neovaginal mucosa where squamous metaplasia didn't occurred. However, In lower neovaginal mucosa where squamous metaplasia occurred, the expression of occludin and ZO-1 in cells were similar to those in control group, which might play a role in the defense function of neovaginal mucosa.

[Different therapeutic efficacy of pralidoxime chloride PAM-Cl on AChE against acute toxicity of methamidophos, dichlorvos and omethoate].

OBJECTIVE: To observe the treatments on the patients with acute methamidophos dichlorvos (DDV) and omethoate poisoning and provide the reliable basis for the rational treatments on these three organophosphorus pesticides poisoning. METHODS: 101 patients with AOPP in 7 hospitals were divided into three groups: Group A, 59 patients with acute methamidophos poisoning, Group B, 32 patients with acute DDV/dipterex (DEP) poisoning, Group C, 10 patients with acute omethoate/dimethoate poisoning. The levels of erythrocyte AChE and the therapeutic efficacies of pralidoxime chloride (PAM-Cl) were compared among the three groups. RESULTS: The AChE activities of all the three groups were inhibited on level of (9.12 +/- 7.99) U/g Hb (group A), 7.32 +/- 4.62 U/g Hb (group B) and (12.01 +/- 9.53) U/g Hb (group C), among which no significant difference was found (P > 0.05). All the patients recovered from acute cholinergic excitation or crisis after the treatment of PAM-Cl. The erythrocyte AChE activities were obviously reactivated in group A three hours later after admission to hospital, each on level of (11.37 +/- 8.67) U/g Hb, (12.51 +/- 6.98) U/g Hb, (15.90 +/- 7.31) U/g Hb, (18.33 +/- 4.78) U/g Hb and (18.91 +/- 7.00) U/g Hb at the 12th, 24th, 48th, 72nd hour and discharge (P < 0.05), and the upgrade tendency was continuous. AChE activities in group B were also reactivated after treatment, each on level of (8.91 +/- 5.89) U/g Hb, (1.31 +/- 6.61) U/g Hb, (13.00 +/- 7.55) U/g Hb, (14.22 +/- 7.80) U/g Hb, (12.78 +/- 7.07) U/g Hb and (16.87 +/- 7.06) U/g Hb at the 3rd, 12th, 24th, 48th, 72nd hour and discharge, but the upgrade tendency turned slowly after 12 hours, the inhibited AChE activities were not reactivated in group C from the beginning to the end. CONCLUSION: After the treatment of PAM-Cl, the AChE activities of the patients with acute methamidophos poisoning could be continuously reactivated, the AChE activities of the patients with acute DDV/DEP poisoning could also be reactivated in 12 hours, and then keep stable, but the AChE activities of the patients with acute omethoate/dimethoate poisoning could not be reactivated. However, PAM-Cl has therapeutic efficacy against acute toxicity of all the three organophosphorus pesticides. Oximes should be vigorously used in the treatment of AOPP, including acute omethoate/dimethoate poisoning.

[Morphologic change and expression of neuronal nitric oxide synthase in neovaginal mucosa of patients undergoing vaginal construction with sigmoid colon].

OBJECTIVE: To study the changes of the histology and expression of neuronal nitric oxide synthase (nNOS) in the neovaginal mucosa of the patients undergoing vaginal construction with sigmoid colon. METHODS: Biopsy samples of upper one-third and lower one-third of the artificial vagina were obtained from 14 patients who underwent vaginal construction with sigmoid colon 9 - 48 months before and samples of normal sigmoid colon were collected from another 14 patients during vaginal construction with sigmoid colon as control group. The pathological and ultrastructural changes were comparatively observed by light microscopy and electron microscopy. At the same time, the expression of nNOS was semiquantitatively evaluated with immunohistochemical method. RESULTS: The upper one-third of the artificial vagina muscular layer of mucosa was hyperplastic and hypertrophic and the density of the submucosal nerve plexus was 3.6 +/- 1.5, significantly higher than that of the control group (1.7 +/- 0.8, P < 0.05). Squamous metaplasia was seen in the colonic mucosa from some 3 samples of lower one-third of the artificial vagina. Partial fusion of the intercellular tight junction, disappearance and fusion of mitochondrial cristae, and degranulation of rough endoplasmic reticulum could be seen in the vaginal colonic mucosa. Compared with the normal control, the expression of nNOS in the vaginal colonic mucosa was significantly decreased. CONCLUSION: Morphologic changes occur nNOS expression decreases in the colonic mucosa of the artificial vagina.