Treatment of Calcified Insertional Achilles Tendinopathy by the Posterior Midline Approach.

The present study investigated the clinical outcomes of the posterior midline approach in the treatment of 34 patients with significantly calcified insertional Achilles tendinopathy. The posterior midline approach was applied for the surgical treatment of 34 patients with chronic significantly calcified insertional Achilles tendinopathy after failed conservative treatment. Gastrocnemius recession was performed simultaneously for patients with gastrocnemius contracture. The Fowler-Philip angle and parallel pitch lines were measured before surgery, and the visual analog scale, Tegner score, and Victorian Institute of Sport tendon study group score were recorded before and after surgery. The mean follow-up period was 45.2 ± 17.7 (range 24 to 84) months. After surgery, the visual analog scale score had decreased notably, and the Tegner score and Victorian Institute of Sport tendon study group score had increased significantly. The posterior midline approach can achieve satisfactory outcomes in the treatment of significantly calcified insertional Achilles tendinopathy, and gastrocnemius recession (Strayer procedure) should be performed for patients with gastrocnemius contracture to improve the surgical outcome.

A new selaginellin derivative from Selaginella pulvinata.

Abstract: A new selaginellin derivative named as selaginellin S (1) was isolated from the whole plants of Selaginella pulvinata (Hook. et Grev.) Maxim. (Selaginellaceae), together with a known one (selaginellin M, 2). Compounds 1 and 2 were separated and purified by silica gel and Sephadex LH-20 column chromatography. Their structures were determined on the basis of extensive spectroscopic analysis including IR, MS, 1D and 2D NMR experiments, as well as ECD calculations. Compound 1 is a key intermidiant in the biosynthesis pathway of selaginellins. Compound 2 is first reported in this plant.

S-nitrosylation of mixed lineage kinase 3 contributes to its activation after cerebral ischemia.

Previous studies in our laboratory have shown that mixed lineage kinase 3 (MLK3) can be activated following global ischemia. In addition, other laboratories have reported that the activation of MLK3 may be linked to the accumulation of free radicals. However, the mechanism of MLK3 activation remains incompletely understood. We report here that MLK3, overexpressed in HEK293 cells, is S-nitrosylated (forming SNO-MLK3) via a reaction with S-nitrosoglutathione, an exogenous nitric oxide (NO) donor, at one critical cysteine residue (Cys-688). We further show that the S-nitrosylation of MLK3 contributes to its dimerization and activation. We also investigated whether the activation of MLK3 is associated with S-nitrosylation following rat brain ischemia/reperfusion. Our results show that the administration of 7-nitroindazole, an inhibitor of neuronal NO synthase (nNOS), or nNOS antisense oligodeoxynucleotides diminished the S-nitrosylation of MLK3 and inhibited its activation induced by cerebral ischemia/reperfusion. In contrast, 2-amino-5,6-dihydro-6-methyl-4H-1,3-thiazine (an inhibitor of inducible NO synthase) or nNOS missense oligodeoxynucleotides did not affect the S-nitrosylation of MLK3. In addition, treatment with sodium nitroprusside (an exogenous NO donor) and S-nitrosoglutathione or MK801, an antagonist of the N-methyl-D-aspartate receptor, also diminished the S-nitrosylation and activation of MLK3 induced by cerebral ischemia/reperfusion. The activation of MLK3 facilitated its downstream protein kinase kinase 4/7 (MKK4/7)-JNK signaling module and both nuclear and non-nuclear apoptosis pathways. These data suggest that the activation of MLK3 during the early stages of ischemia/reperfusion is modulated by S-nitrosylation and provides a potential new approach for stroke therapy whereby the post-translational modification machinery is targeted.

Research progress in the mechanism of effect of PRP in bone deficiency healing.

Platelet-rich plasma (PRP) therapy is a recently developed technique that uses a concentrated portion of autologous blood to try to improve and accelerate the healing of various tissues. There is a considerable interest in using these PRP products for the treatment used in bone deficiency healing. Because PRP products are safe and easy to prepare and administer, there has been increased attention toward using PRP in numerous clinical settings. The benefits of PRP therapy appear to be promising, and many investigators are exploring the ways in which this therapy can be used in the clinical setting. At present, the molecular mechanisms of bone defect repair studies have focused on three aspects of the inflammatory cytokines, growth factors and angiogenic factors. The role of PRP works mainly through these three aspects of bone repair. The purpose of this paper is to review the current evidence on the mechanism of the effect of PRP in bone deficiency healing.

Forecast for peak infections in the second wave of the Omicron after the adjustment of zero-COVID policy in the mainland of China.

On December 7, 2022, the Chinese government optimized the current epidemic prevention and control policy, and no longer adopted the zero-COVID policy and mandatory quarantine measures. Based on the above policy changes, this paper establishes a compartment dynamics model considering age distribution, home isolation and vaccinations. Parameter estimation was performed using improved least squares and Nelder-Mead simplex algorithms combined with modified case data. Then, using the estimated parameter values to predict a second wave of the outbreak, the peak of severe cases will reach on 8 May 2023, the number of severe cases will reach 206,000. Next, it is proposed that with the extension of the effective time of antibodies obtained after infection, the peak of severe cases in the second wave of the epidemic will be delayed, and the final scale of the disease will be reduced. When the effectiveness of antibodies is 6 months, the severe cases of the second wave will peak on July 5, 2023, the number of severe cases is 194,000. Finally, the importance of vaccination rates is demonstrated, when the vaccination rate of susceptible people under 60 years old reaches 98%, and the vaccination rate of susceptible people over 60 years old reaches 96%, the peak of severe cases in the second wave of the epidemic will be reached on 13 July 2023, when the number of severe cases is 166,000.

Glioblastoma cellular MAP4K1 facilitates tumor growth and disrupts T effector cell infiltration.

MAP4K1 has been identified as a cancer immunotherapy target. Whether and how cancer cell-intrinsic MAP4K1 contributes to glioblastoma multiforme (GBM) progression remains unclear. We found that MAP4K1 was highly expressed in the glioma cells of human GBM specimens. High levels of MAP4K1 mRNA were prevalent in IDH-WT and 1p/19q non-codeletion gliomas and correlated with poor prognosis of patients. MAP4K1 silencing inhibited GBM cell proliferation and glioma growth. Transcriptome analysis of GBM cells and patient samples showed that MAP4K1 modulated cytokine‒cytokine receptor interactions and chemokine signaling pathway, including IL-18R and IL-6R Importantly, MAP4K1 loss down-regulated membrane-bound IL-18R/IL-6R by inhibiting the PI3K-AKT pathway, whereas MAP4K1 restoration rescued this phenotype and therefore GBM cell proliferation. MAP4K1 deficiency abolished GBM cell pro-proliferation responses to IL-18, suggesting an oncogenic role of MAP4K1 via the intrinsic IL-18/IL-18R pathway. In addition, GBM cell-derived MAP4K1 impaired T-cell migration and reduced CD8+ T-cell infiltration in mouse glioma models. Together, our findings provide novel insight into the pathological significance of GBM cell-intrinsic MAP4K1 in driving tumor growth and immune evasion by remodeling cytokine-chemokine networks.

Activation of GluR6-containing kainate receptors induces ubiquitin-dependent Bcl-2 degradation via denitrosylation in the rat hippocampus after kainate treatment.

We previously showed that Bcl-2 (B-cell lymphoma 2) is down-regulated in a kainate (KA)-induced rat epileptic seizure model. The underlying mechanism had remained largely unknown, but we here report for the first time that denitrosylation and ubiquitination are involved. Our results show that the S-nitrosylation levels of Bcl-2 are down-regulated after KA injection and that the GluR6 (glutamate receptor 6) antagonist NS102 can inhibit the denitrosylation of Bcl-2. Moreover, the ubiquitin-dependent degradation of Bcl-2 was found to be promoted after KA treatment, which could be suppressed by the proteasome inhibitor MG132 and the NO donors, sodium nitroprusside and S-nitrosoglutathione. In addition, experiments based on siRNA transfections were performed in the human SH-SY5Y neuroblastoma cell line to verify that the stability of Bcl-2 is causal to neuronal survival. At the same time, it was found that the exogenous NO donor GSNO could protect neurons when Bcl-2 is targeted. Subsequently, these mechanisms were morphologically validated by immunohistochemistry, cresyl violet staining, and in situ TUNEL staining to analyze the expression of Bcl-2 as well as the survival of CA1 and CA3/DG pyramidal neurons. NS102, GSNO, sodium nitroprusside, and MG132 contribute to the survival of CA1 and CA3/DG pyramidal neurons by attenuating Bcl-2 denitrosylation. Taken together, our data reveal that Bcl-2 ubiquitin-dependent degradation is induced by Bcl-2 denitrosylation during neuronal apoptosis after KA treatment.

S-nitrosylation of c-Src via NMDAR-nNOS module promotes c-Src activation and NR2A phosphorylation in cerebral ischemia/reperfusion.

Previous studies suggested that activated c-Src promote the tyrosine phosphorylation of NMDA receptor subunit NR2A, and thus aggravate the injury induced by transient cerebral ischemia/reperfusion (I/R) in rat hippocampus CA1 region. In this study, we examined the effect of nitric oxide (NO) on the activation of c-Src and the tyrosine phosphorylation of NMDA receptor NR2A subunit. The results show that S-nitrosylation and the phosphorylation of c-Src were induced after cerebral I/R in rats, and administration of nNOS inhibitor 7-NI, nNOS antisense oligonucleotides and exogenous NO donor sodium nitroprusside diminished the increased S-nitrosylation and phosphorylation of c-Src during cerebral I/R. The cysteine residues of c-Src modified by S-nitrosylation are Cys489, Cys498, and Cys500. On the other hand, NMDAR antagonist MK-801 could attenuate the S-nitrosylation and activation of c-Src. Taken together, the S-nitrosylation of c-Src is provoked by NO derived from endogenous nNOS, which is activated by Ca(2+) influx from NMDA receptors, and promotes the auto-phosphorylation at tyrosines and further phosphorylates NR2A. The molecular mechanism we outlined here is a novel postsynaptic NMDAR-nNOS/c-Src-mediated signaling amplification, the 'NMDAR-nNOS â†’ NO â†’ SNO-c-Src â†’ p-c-Src â†’ NMDAR-nNOS' cycle, which presents the possibility as a potential therapeutic target for stroke treatment.

Cerebral ischemia-reperfusion induces GAPDH S-nitrosylation and nuclear translocation.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a glycolytic enzyme, plays an important role in glycolysis. It was reported that GAPDH undergoes S-nitrosylation, which facilitated its binding to Siah1 and resulted in nuclear translocation and cell apoptosis. The results of this study show that GAPDH S-nitrosylation, Siah1 binding, translocation to nucleus, and concomitant neuron death occur during the early stages of reperfusion in the rat four-vessel occlusion ischemic model. N-Methyl-D-aspartate receptor antagonist MK801, neuronal nitric oxide synthase inhibitor 7-nitroindazole, or monoamine oxidase-B inhibitor (R)-(-)-deprenyl hydrochloride could inhibit GAPDH S-nitrosylation and translocation and exert neuroprotective effects.

Inflammation & apoptosis in spinal cord injury.

Spinal cord injury (SCI) consists of a two-steps process involving a primary mechanical injury followed by an inflammatory process and apoptosis. Secondary insult is characterized by further destruction of neuronal and glial cells, and leads to expansion of the damage, so that the paralysis can extend to higher segments. With the identification of mechanisms that either promote or prevent neuronal inflammation and apoptosis come new approaches for preventing and treating neurodegenerative disorders. From a clinical perspective, this article discusses novel targets for the development of therapeutic agents that have the potential to protect the spinal cord from irreversible damage and promote functional recovery.

Expression, purification, and characterization of RGD-mda-7, a His-tagged mda-7/IL-24 mutant protein.

RGD peptide (Arg-Gly-Asp tripeptide) binds to integrin αVß(3) and αVß(5), which is selectively expressed in tumor neovasculature and on the surface of some tumor cells. Some studies showed that coupling the RGD peptides to anticancer drugs yielded compounds with increased efficiency against tumors and lowered toxicity to normal tissues. The melanoma differentiation-associated gene-7/interleukin-24 gene (mda-7/IL-24) is a novel tumor-suppressor/cytokine gene that exhibits potent tumor-suppressive activity without damaging normal cells. To enhance the antitumor effect, we inserted a glycine residue into the wild type (mda-7/IL-24) between (164)Arg and (165)Asp to form a RGD peptide, named RGD-mda-7, then expressed RGD-mda-7 in Escherichia coli. Herein, we describe the expression and purification of RGD-mda-7. We detected the characterizations of immunostimulatory activity, tumor targeting, potent cytopathic effect, and apoptosis inducing exploited by RGD-mda-7 in tumor cells, and also compared these characterizations with wtmda-7/IL-24. The data showed that RGD-mda-7 had more potent tumor targeting and apoptosis-inducing effects than wtmda-7/IL-24.

Impacts of climate change on vegetation pattern: Mathematical modeling and data analysis.

Climate change has become increasingly severe, threatening ecosystem stability and, in particular, biodiversity. As a typical indicator of ecosystem evolution, vegetation growth is inevitably affected by climate change, and therefore has a great potential to provide valuable information for addressing such ecosystem problems. However, the impacts of climate change on vegetation growth, especially the spatial and temporal distribution of vegetation, are still lacking of comprehensive exposition. To this end, this review systematically reveals the influences of climate change on vegetation dynamics in both time and space by dynamical modeling the interactions of meteorological elements and vegetation growth. Moreover, we characterize the long-term evolution trend of vegetation growth under climate change in some typical regions based on data analysis. This work is expected to lay a necessary foundation for systematically revealing the coupling effect of climate change on the ecosystem.

Antimicrobial selaginellin derivatives from Selaginella pulvinata.

Six selaginellin derivatives, including three new analogues selaginellins D-F (1-3), were isolated from the EtOAc extract of the whole plant of Selaginella pulvinata (Hook. et Grev.) Maxim. Their structures were determined on the basis of extensive physical and chemical evidence. Compounds 1 and 4 demonstrated antifungal activities against Candida albicans; compounds 4-6 exhibited significant antibacterial activity against Staphylococcus aureus.

Structure determination of selaginellins G and H from Selaginella pulvinata by NMR spectroscopy.

Selaginellins G (1) and H (2), two new selaginellin derivatives, were isolated from the whole plant of Selaginella pulvinata. Their structures were elucidated, and complete assignments of the (1)H and (13)C NMR spectroscopic data were achieved by 1D and 2D NMR experiments (HSQC, HMBC, COSY and ROESY). Compound 1 displayed good antifungal activity against Candida albicans with an IC(50) value of 5.3 microg/ml.

MLK3 Is Associated With Poor Prognosis in Patients With Glioblastomas and Actin Cytoskeleton Remodeling in Glioblastoma Cells.

Mixed lineage kinase 3 (MLK3) has been implicated in human melanoma and breast cancers. However, the clinical significance of MLK3 in human gliomas and the underlying cellular and molecular mechanisms remain unclear. We found that MLK3 proteins were highly expressed in high-grade human glioma specimens and especially prevalent in primary and recurrent glioblastoma multiforme (GBM). High levels of MLK3 mRNA were correlated with poor prognosis in patients with isocitrate dehydrogenase (IDH)-wild-type (wt) gliomas. Furthermore, genetic ablation of MLK3 significantly suppressed the migration and invasion abilities of GBM cells and disrupted actin cytoskeleton organization. Importantly, MLK3 directly bound to epidermal growth factor receptor kinase substrate 8 (EPS8) and regulated the cellular location of EPS8, which is essential for actin cytoskeleton rearrangement. Overall, these findings provide evidence that MLK3 upregulation predicts progression and poor prognosis in human IDH-wt gliomas and suggest that MLK3 promotes the migration and invasion of GBM cells by remodeling the actin cytoskeleton via MLK3-EPS8 signaling.

An in vivo mouse model of long-term potentiation at synapses between primary afferent C-fibers and spinal dorsal horn neurons: essential role of EphB1 receptor.

BACKGROUND: Long-term potentiation (LTP), a much studied cellular model of synaptic plasticity, has not been demonstrated at synapses between primary afferent C-fibers and spinal dorsal horn (DH) neurons in mice in vivo. EphrinB-EphB receptor signaling plays important roles in synaptic connection and plasticity in the nervous system, but its role in spinal synaptic plasticity remains unclear. RESULTS: This study characterizes properties of LTP at synapses of C-fibers onto neurons in the superficial DH following high-frequency stimulation (HFS) of a peripheral nerve at an intensity that activates C-fibers and examines associated activation of Ca2+/calmodulin-activated protein kinase II (p-CaMKII), extracellular signal-regulated kinase (p-ERK) and the cyclic AMP response element binding protein (p-CREB) and expression of c-Fos, and it investigates further roles for the EphB1 receptor in LTP. HFS induced LTP within 5 min and lasts for 3-8 h during the period of recording and resulted in upregulation of p-CaMKII, p-ERK and p-CREB protein levels in the spinal cord and expression of c-Fos in DH. Intrathecal pretreatment of MK-801 or EphB2-Fc prevented LTP and significantly reduced upregulation of p-CaMKII, p-ERK, p-CREB and c-Fos. Further, targeted mutation of EphB1 receptor prevented induction of LTP and associated increases in phosphorylation of CaMKII, ERK, and CREB. CONCLUSION: This study provides an in vivo mouse model of LTP at synapses of C-fibers onto the superficial DH neurons that will be valuable for studying the DH neuron excitability and their synaptic plasticity and hyperalgesia. It further takes advantage of examining functional implications of a specific gene targeted mice and demonstrates that the EphB1 receptor is essential for development of LTP.

[Effect of arsenic trioxide combined with aspirin on the growth of human gastric adenocarcinoma graft in nude mice].

OBJECTIVE: To investigate the inhibitory effect of arsenic trioxide, aspirin and their combination on the growth of human gastric adenocarcinoma SGC-7901 graft in nude mice. METHODS: Thirty-one model nude mice with transplanted tumor of human gastric adenocarcinoma SGC-7901 were established successfully by fixing tumor tissue under the left axillary subcutaneously. They were randomly divided into 4 groups and treated respectively with normal saline (control group, n = 7), aspirin (Asp group, n = 8), arsenic trioxide (AT group, n = 8), and AT +Asp (combined treated group, n = 8), medicated by intraperitoneal injection for 14 successive days. All mice were sacrificed 48 h after the final medication, the transplanted tumor mass volume and weight were measured to calculate the tumor inhibition rate, and changes of tumors and tissues of heart, liver, and kidney, etc were observed. Moreover, expressions of proliferation associated protein Ki67 and apoptosis inhibitory protein Bcl-2 were detected by immunohistochemical SP method, and expression of apoptosis associated protein Bax was detected by Western blotting. RESULTS: After terminating the medication, in the combined treated group the tumor volume and weight were significantly lower than those in the control group (P < 0.05) and the tumor weight was lower than that in the AT and Asp groups (both P < 0.05). The volume inhibition rate was 59.12% in the AT group, 47.60% in the Asp group, and 76.70% in the combined treated group, and the weight inhibition rate in them 35.60%, 45.44% and 63.90%, respectively. Immunochemical staining showed that as compared with the control group, expressions of Ki67 and Bcl-2 were down-regulated in all the three treated groups to some extent (all P < 0.05), and significant difference was also shown in comparison of the two indices between the combined treated group with the other two treated groups (P < 0.05). The expression of Bax was higher in the combined treated group than in other groups (all P < 0.05). CONCLUSION: Combined use of AT and Asp has certain cooperative effect in inhibiting the growth of gastric carcinoma cells without obvious toxic-side effects.

[Warm acupuncture on chronic obstructive pulmonary disease with phlegm-turbid obstructing of the lung: a randomized controlled trial].

OBJECTIVE: To verify the effect of warm acupuncture combined with western medicine in patients with acute exacerbation chronic obstructive pulmonary disease (AECOPD) with phlegm-turbid obstructing of the lung. METHODS: Ninety patients with AECOPD were randomly divided into a western medicine group, a warm acupuncture group and a sham acupuncture group, 30 cases in each group. Routine treatment according to the Global initiative for chronic obstructive lung disease (GOLD) guidelines was used in the western medicine group. On the basis of the routine treatment, the warm acupuncture was applied at Fenglong (ST 40), Feishu (BL 13), Zusanli (ST 36) and other acupoints in the warm acupuncture group. In the sham acupuncture group, non-acupoints were taken on the basis of conventional treatment, and superficial acupuncture was performed for 1-3 mm, once a day for 2 weeks. Forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC), FEV1/FVC, cough and sputum assessment questionnaire (CASA), COPD assessment test (CAT), and the change of TCM syndrome score were observed before and after treatment in all groups. RESULTS: After treatment, the FEV1, FVC and FEV1/FVC in each group were higher than those before treatment (P<0.05), there was no significant difference among groups after treatment (P>0.05). The CASA score, CAT score and TCM syndrome score in each group were lower than those before treatment (P<0.05). The change of the scores before and after treatment in the warm acupuncture group was higher than that in the sham acupuncture group and the western medicine group (P<0.05). There was no significant difference between the sham acupuncture group and the western medicine group (P>0.05). CONCLUSION: Warm acupuncture has a significant effect on the improvement of symptoms in patients with AECOPD with phlegm-turbid obstructing of the lung.

Therapeutic Outcomes of Kalix II in Treating Juvenile Flexible Flatfoot.

OBJECTIVES: To evaluate the therapeutic outcomes with Kalix II subtalar arthroereisis in sinus tarsi for juvenile flexible flatfoot. METHODS: A retrospective analysis of the data of 20 juveniles with symptomatic flexible flatfoot (27 feet) who underwent the Kalix II implant procedure from January 2008 to September 2012 was performed. The pain during daily activities was assessed and followed up by use of a standard 10-point visual analog scale (VAS), and function was evaluated using the American Orthopaedic Foot and Ankle Society (AOFAS) ankle and hindfoot scoring system, and anteroposterior talar-first metatarsal angle, lateral talar-first metatarsal angle, calcaneal pitch angle, and talar declination angle at X-ray film were measured to assess the therapeutic outcomes. Patients were asked to grade the result of the procedure as excellent, good, fair, or poor at latest follow-up. The data was expressed as mean ± standard deviation (SD). A paired Student's t -test was used for comparisons of the preoperative and postoperative angular measurements for each foot, VAS scores, and AOFAS scores. In all tests, P < 0.05 was considered statistically significant. RESULTS: The mean age of the patients was 12.1 years (range, 7-16 years), and 16 left feet and 11 right feet were involved. All patients finished the follow-up with a mean period of 28.1 months (range, 23-60 months). Eleven feet were treated with subtalar arthroereisis combined with reconstruction of the end point of the posterior tibialis tendon after dissection of the accessory scaphoid. The subtalar arthroereisis device displaced in 1 foot due to a fall from the inversion position 3 months after surgery, and was replaced by a new device after the failure of conservative treatment. The mean VAS score decreased from 5.6 ± 0.5 preoperatively to 1.2 ± 0.2 (P < 0.001), and the mean AOFAS hindfoot and ankle score improved from 71.1 ± 6.1 preoperatively to 88.1 ± 6.3 (P < 0.001). Differences between preoperative and postoperative measurements for each radiographic variable were statistically significant (P < 0.001). Comparison of radiographic measurements showed that the anteroposterior talar-first metatarsal (Meary) angle decreased by a mean of 12.8° ± 1.5°, the lateral talar-first metatarsal (Meary) angle decreased by a mean of 15.4° ± 1.3°, the calcaneal pitch angle increased by a mean of -2.1° ± 0.7°, and the talar declination angle decreased by a mean of 17.9° ± 2.8°. Overall, 12 patients rated the result as excellent, 6 as good, and 2 as fair. CONCLUSION: The application of Kalix II in subtalar arthroereisis combined with dissection of accessory scaphoid and reconstruction of posterior tibialis tendon is an effective therapy for flexible juvenile flatfoot.