MMP9 Differentially Regulates Proteins Involved in Actin Polymerization and Cell Migration during TGF-ß-Induced EMT in the Lens.

Fibrotic cataracts have been attributed to transforming growth factor-beta (TGF-ß)-induced epithelial-to-mesenchymal transition (EMT). Using mouse knockout (KO) models, our laboratory has identified MMP9 as a crucial protein in the TGF-ß-induced EMT process. In this study, we further revealed an absence of alpha-smooth muscle actin (αSMA) and filamentous-actin (F-actin) stress fibers in MMP9KO mouse lens epithelial cell explants (LECs). Expression analysis using NanoString revealed no marked differences in αSMA (ACTA2) and beta-actin (ß-actin) (ACTB) mRNA between the lenses of TGF-ß-overexpressing (TGF-ßtg) mice and TGF-ßtg mice on a MMP9KO background. We subsequently conducted a protein array that revealed differential regulation of proteins known to be involved in actin polymerization and cell migration in TGF-ß-treated MMP9KO mouse LECs when compared to untreated controls. Immunofluorescence analyses using rat LECs and the novel MMP9-specific inhibitor, JNJ0966, revealed similar differential regulation of cortactin, FAK, LIMK1 and MLC2 as observed in the array. Finally, a reduction in the nuclear localization of MRTF-A, a master regulator of cytoskeletal remodeling during EMT, was observed in rat LECs co-treated with JNJ0966 and TGF-ß. In conclusion, MMP9 deficiency results in differential regulation of proteins involved in actin polymerization and cell migration, and this in turn prevents TGF-ß-induced EMT in the lens.

Chemical Composition, Antioxidative and Anticancer Activities of the Essential Oil: Curcumae Rhizoma-Sparganii Rhizoma, a Traditional Herb Pair.

As a classical herb pair in clinics of traditional Chinese medicine, Curcumae Rhizoma-Sparganii Rhizoma (HP CR-SR) is used for activating blood circulation to remove blood stasis. The essential components in HP CR-SR and its single herbs were comparatively analyzed using gas chromatography-mass spectrometry data. 66, 22, and 54 components in volatile oils of Curcumae Rhizoma, Sparganii Rhizoma, and HP CR-SR were identified, and total contents accounted for 75.416%, 91.857%, and 79.553% respectively. The thirty-eight components were found in HP CR-SR, and not detected in single herbs Curcumae Rhizoma and Sparganii Rhizoma. The highest radical trapping action was seen by an essential oil of HP CR-SR (IC50 = 0.59 ± 0.04 mg/mL). Furthermore, the HP CR-SR essential oil showed more remarkable cytotoxicity on tumor cell lines than that of the single herbs Curcumae Rhizoma and Sparganii Rhizoma in a dose-dependent manner: IC50 values showing 32.32 ± 5.31 µg/mL (HeLa), 34.76 ± 1.82 µg/mL (BGC823), 74.84 ± 1.66 µg/mL (MCF-7), 66.12 ± 11.23 µg/mL (SKOV3), and 708.24 ± 943.91 µg/mL (A549), respectively. In summary, the essential oil of HP CR-SR is different from any one of Curcumae Rhizoma and Sparganii Rhizoma, nor simply their superposition, and HP CR-SR oil presented more remarkable anticancer and antioxidant activities compared with Curcumae Rhizoma and Sparganii Rhizoma oils.

Spectrum-effect relationships as a systematic approach to traditional chinese medicine research: current status and future perspectives.

Component fingerprints are a recognized method used worldwide to evaluate the quality of traditional Chinese medicines (TCMs). To foster the strengths and circumvent the weaknesses of the fingerprint technique in TCM, spectrum-effect relationships would complementarily clarify the nature of pharmacodynamic effects in the practice of TCM. The application of the spectrum-effect relationship method is crucial for understanding and interpreting TCM development, especially in the view of the trends towards TCM modernization and standardization. The basic requirement for using this method is in-depth knowledge of the active material basis and mechanisms of action. It is a novel and effective approach to study TCMs and great progress has been made, but to make it more accurate for TCM research purposes, more efforts are needed. In this review, the authors summarize the current knowledge about the spectrum-effect relationship method, including the fingerprint methods, pharmacodynamics studies and the methods of establishing relationships between the fingerprints and pharmacodynamics. Some speculation regarding future perspectives for spectrum-effect relationship approaches in TCM modernization and standardization are also proposed.

Gaultheria: Phytochemical and pharmacological characteristics.

The genus Gaultheria, comprised of approximately 134 species, is mostly used in ethnic drugs to cure rheumatism and relieve pain. Phytochemical investigations of the genus Gaultheria have revealed the presence of methyl salicylate derivatives, C6-C3 constituents, organic acids, terpenoids, steroids, and other compounds. Methyl salicylate glycoside is considered as a characteristic ingredient in this genus, whose anti-rheumatic effects may have a new mechanism of action. In this review, comprehensive information on the phytochemistry, volatile components and the pharmacology of the genus Gaultheria is provided to explore its potential and advance research.

Electrophysiological mechanisms of motion-style scalp acupuncture for treating poststroke spasticity in rats. / è¿åŠ¨å¤´é’ˆæ³•æŠ—å’ä¸­åŽç—‰æŒ›æ•ˆåº”çš„ç”µç”Ÿç†æœºåˆ¶ç ”ç©¶.

OBJECTIVES: To observe the effect of motion-style scalp acupuncture (MSSA) on H-reflex in rats with post-stroke spasticity (PSS), so as to explore the electrophysiological mechanisms of MSSA against spasticity. METHODS: A total of 36 male SD rats were randomly divided into sham operation, model and MSSA groups, with 12 rats in each group. The stroke model was established by occlusion of the middle cerebral artery. After modeling, rats in the MSSA group were treated by scalp acupuncture (manipulated every 15 min, 200 r/min) at ipsilesional "parietal and temporal anterior oblique line" (MS6) for a total of 30 min, the treadmill training (10 m/min) was applied during the needling retention, once daily for consecutive 7 days. The neurological deficits, muscle tone and motor function were assessed by Zea Longa score, modified modified Ashworth scale (MMAS) score and screen test score before and after treatment, respectively. The H-reflex of spastic muscle was recorded by electrophysiological recordings and the frequency dependent depression (FDD) of H-reflex was also recorded. The cerebral infarction volume was evaluated by TTC staining. RESULTS: Compared with the sham operation group, the Zea longa score, MMAS score, cerebral infarction volume, motion threshold, Hmax/Mmax ratio and FDD of H-reflex were significantly increased (P<0.01), while the screen test score was significantly decreased (P<0.01) in the model group. Intriguingly, compared with the model group, the above results were all reversed (P<0.01) in the MSSA group. CONCLUSIONS: MSSA could exert satisfactory anti-spastic effects in rats with PSS, the underlying mechanism may be related to the improvement of nerve function injury, the reduction of spastic muscle movement threshold, Hmax/Mmax ratio and H-reflex FDD.

m6A Modification Patterns With Distinct Immunity, Metabolism, and Stemness Characteristics in Soft Tissue Sarcoma.

N6-methyladenosine (m6A) RNA methylation has been shown to have prognostic value in cancer. Nonetheless, its potential role regarding immunity, metabolism, and stemness in soft tissue sarcoma (STS) remains unknown. We comprehensively estimated the m6A modification patterns and corresponding immunity, metabolism, and stemness characteristics based on 568 STS samples and 21 m6A regulators. The m6Ascore was constructed to quantify m6A modification patterns in individuals using machine learning algorithms. Two distinct m6A modification patterns among the STS patients were identified, which exhibited differences in prognosis, immune cell infiltration, metabolic pathways, stemness, somatic mutation, and copy number variation. Thereafter, immunity-, metabolism-, and stemness phenotype-related genes associated with m6A modification were identified. Furthermore, patients with lower m6Ascores had increased antitumor immune responses, survival benefit under immunotherapy, tumor mutation burden, immunogenicity, and response to anti-PD-1/L1 immunotherapy. Immunotherapy sensitivity was validated using the IMvigor210 dataset. STS patients with lower m6Ascore might be more sensitive to docetaxel and gemcitabine. Finally, pan-cancer analysis illustrated the significant correlations of m6Ascore with clinical outcomes, immune cell infiltration, metabolism, and stemness. This study revealed that m6A modification plays an important role in immunity, metabolism, and stemness in STS. Evaluating the m6A modification pattern and development of m6Ascore may help to guide more effective immunotherapy and chemotherapy strategies.

Molecular Subtypes Based on Cell Differentiation Trajectories in Head and Neck Squamous Cell Carcinoma: Differential Prognosis and Immunotherapeutic Responses.

Objective: Head and neck squamous cell carcinoma (HNSCC) is one of the most common and lethal malignant tumors. We aimed to investigate the HNSCC cell differentiation trajectories and the corresponding clinical relevance. Methods: Based on HNSCC cell differentiation-related genes (HDRGs) identified by single-cell sequencing analysis, the molecular subtypes and corresponding immunity, metabolism, and stemness characteristics of 866 HNSCC cases were comprehensively analyzed. Machine-learning strategies were used to develop a HNSCC cell differentiation score (HCDscore) in order to quantify the unique heterogeneity of individual samples. We also assessed the prognostic value and biological characteristics of HCDscore using the multi-omics data. Results: HNSCCs were stratified into three distinct molecular subtypes based on HDRGs: active stroma (Cluster-A), active metabolism (Cluster-B), and active immune (Cluster-C) types. The three molecular subtypes had different characteristics in terms of biological phenotype, genome and epigenetics, prognosis, immunotherapy and chemotherapy responses. We then demonstrated the correlations between HCDscore and the immune microenvironment, subtypes, carcinogenic biological processes, genetic variation, and prognosis. The low-HCDscore group was characterized by activation of immunity, enhanced response to anti-PD-1/PD-L1 immunotherapy, and better survival compared to the high-HCDscore group. Finally, by integrating the HCDscore with prognostic clinicopathological characteristics, a nomogram with strong predictive performance and high accuracy was constructed. Conclusions: This study revealed that the cell differentiation trajectories in HNSCC played a nonnegligible role in patient prognosis, biological characteristics, and immune responses. Evaluating cancer cell differentiation will help to develop more effective immunotherapy, metabolic therapy, and chemotherapy strategies.

Integrated Analysis of Nine Prognostic RNA-Binding Proteins in Soft Tissue Sarcoma.

RNA-binding proteins (RBPs) have been shown to be dysregulated in cancer transcription and translation, but few studies have investigated their mechanism of action in soft tissue sarcoma (STS). Here, The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases were used to identify differentially expressed RBPs in STS and normal tissues. Through a series of biological information analyses, 329 differentially expressed RBPs were identified. Functional enrichment analysis showed that differentially expressed RBPs were mainly involved in RNA transport, RNA splicing, mRNA monitoring pathways, ribosome biogenesis and translation regulation. Through Cox regression analyses, 9 RBPs (BYSL, IGF2BP3, DNMT3B, TERT, CD3EAP, SRSF12, TLR7, TRIM21 and MEX3A) were all up-regulated in STS as prognosis-related genes, and a prognostic model was established. The model calculated a risk score based on the expression of 9 hub RBPs. The risk score could be used for risk stratification of patients and had a high prognostic value based on the receiver operating characteristic (ROC) curve. We also established a nomogram containing risk scores and 9 key RBPs to predict the 1-year, 3-year, and 5-year survival rates of patients in STS. Afterwards, methylation analysis showed significant changes in the methylation degree of BYSL, CD3EAP and MEX2A. Furthermore, the expression of 9 hub RBPs was closely related to immune infiltration rather than tumor purity. Based on the above studies, these findings may provide new insights into the pathogenesis of STS and will provide candidate biomarkers for the prognosis of STS.

[Closed reduction and internal fixation versus transtarsal sinus small incision internal fixation for Sanders type II calcaneal fractures].

OBJECTIVE: To compare the clinical effects of closed reduction and internal fixation with limited transtarsal sinus incision in the treatment of Sanders type II calcaneal fractures. METHODS: From June 2015 to February 2018, 60 patients with Sanders type II calcaneal fractures were treated by minimally invasive method. According to the different treatment methods, they were divided into two groups: closed prying reduction and internal fixation with hollow nails(group A) and limited transtarsal sinus incision internal fixation (group B). There were 34 cases in group A, including 22 males and 12 females, aged 21 to 60(38.90±3.25) years old, 14 cases of Sanders type IIA, 14 cases of Sanders type IIB and 6 cases of Sanders type IIC. There were 20 cases in group B, including 14 males and 6 females, aged 20 to 59(39.20±2.96) years old, 8 cases of Sanders type IIA, 9 cases of Sanders type IIB and 3 cases of Sanders type IIC. Calcaneal height, calcaneal width, Gissane angle and Bohler angle were measured before and after operation. The complications, hospitalization time and cost were compared. Functional evaluation was performed by AOFAS score system one year after operation. RESULTS: Fifty-four cases were followed up for 6 to 16(11.40±2.55) months and 6 cases were lost. All the wounds healed in grade A without complications such as skin infection and necrosis. There were 1 case of nerve injury in group A, 2 cases of tendon entrapment, 2 cases of nerve injury in group B, there was no significant difference(P>0.05). There was no significant difference in calcaneal width, height, Bohlers angle and Gissane angle between group A and group B (P>0.05). There was no significant difference in AOFAS score between group A and group B(89.2±4.7 vs 88.7±4.8, P>0.05). The hospitalization expenditure and length of stay in group A were(5 021.25±1 047.19) yuan, (6.31±4.75) d, and those in group B were(13 591.35±1 975.21) yuan, (9.65±3.42) d, respectively, with significant difference(P<0.05). CONCLUSIONS: Closed reduction and internal fixation is as effective as transtarsal sinus incision reduction and internal fixation for Sanders typeIIfracture. It has the advantages of less trauma and fewer complications. Closed reduction and hollow nail fixation group can significantly shorten hospitalization time, reduce hospitalization costs, and is more economical. It is worth recommending, but attention should be paid to avoid choosing to comminute serious calcaneal fracture.

[Clinical trials of acupuncture treatment of post-stroke dysphagia by deep acupuncture of Tiantu (CV22) in combination with swallowing rehabilitation training].

OBJECTIVE: To observe the effect of deep acupuncture of Tiantu (CV22), etc. in combination with rehabilitation training on swallowing function of patients with post-stroke dysphasia. METHODS: A total of 70 patients suffering from post-stroke dysphagia were randomly divided into control group (conventional acupuncture＋ swallowing rehabilitation training) and treatment group (deep acupuncture of CV22 ＋ rehabilitation training, n＝ 35 cases in each group). The swallowing rehabilitation procedure was conducted 30 min every time, twice daily, 5 times a week for 3 weeks. In patients of the treatment group, after deep insertion (about 80 mm) along the posterior margin of the manubrium sterna, the filiform needle in CV22 was lifted and thrusted repeatedly 9 times till the patient experienced a feeling of foreign matter blocking, followed by retaining the needle for 30 min. For patients of the control group, Lianquan (CV23) was punctured with 3 acupuncture needles (one vertical needling and bilateral slope needling toward the tongue root), and Fengfu (GV16), bilateral Bailao (EX-NH23), bilateral Renying (ST9) and bilateral Jialianquan (Extra) were punctured vertically with filiform needles. The treatment was conducted once daily, 5 times a week for 3 weeks. The swallowing function was evaluated by using Kubota water swallowing test (KWST) scale (0－5 points), Fujishima Ichiro rating scale (FIRS, 0－10 points), and video fluoroscopic swallowing study (VFSS, 0－10 points), separately. The therapeutic effect was also assessed according to the clinical scale scores and VFSS results. RESULTS: Following the treatment, of the two 35 cases in the control and treatment groups, 4 (11.43%) and 8 (22.86%) experienced marked improvement in their symptoms, 22 (62.86%) and 23 (65.71%) were effective, and 9 (25.71%) and 4 (11.43%) were failed, with the effective rates being 74.29% and 88.57%, respectively. After the treatment, the KWST score of the two groups were significantly decreased in comparison with their own pre-treatment (P<0.05), while the FIRS and VFSS scores of the two groups were considerably increased in comparison with their own pre-treatment (P<0.05). The therapeutic effects of the treatment group were obviously superior to those of the control group in the effective rate, lowering KWST score and in raising FIRS and VFSS scores (P<0.05)．. CONCLUSION: Acupuncture plus swallowing rehabilitation training is effective in improving the swallowing function of stroke patients with dysphagia, and the deep acupuncture stimulation of CV22 is apparently better than conventional acupuncture in the therapeutic effect.

[Combined posterior and anterior approaches for the treatment of cervical spondylotic myelopathy].

OBJECTIVE: To explore the efficacy of one stage or staging treatment of cervical spondylotic myelopathy through combined posterior and anterior approaches. METHODS: From June 2001 to October 2008, 45 patients with pinching cervical spondylotic myelopathy and developmental stenosis of cervical spinal canal were decompressed through combined posterior and anterior approaches in one stage or staging operation. Among the patients, 35 patients were male and 10 patients were female, ranging in age from 45 to 72 years, with an average of 53 years. Five patients were decompressed through combined posterior and anterior approaches in one-stage operation, and other 40 patients were treated in staging operation. All the patients were treated with open-door posterior cervical expansive laminoplasty and anterior decompression, bone graft and titanium plate internal fixation. JOA scores were used to evaluate the therapeutic effects at the 3rd month and one year after operation. RESULTS: After the treatment, 44 patients were followed up from 13 to 62 months, with an average of 30 months. One patient was dead of lung infection at the 21th day after operation. The nerve root palsy of C5 was found in 3 cases. One patient had postoperative hoarseness. Axial symptoms were found in 14 cases. Two patients had donor site complications. There were no screws broken or back out, no screw loosening, no moving or sinking of bone block or Ti-net, no door re-closer and cervical deformity. Forty-four patients got spinal fusion. According to JOA score criteria: the average preoperative JOA score was (9.36 +/- 2.24) points, JOA score was (12.34 +/- 2.64) points in 3 months after operation, (12.77 +/- 2.61) points in one year after operation. Among 44 cases, 16 got an excellent result, 19 good, 6 fair, 3 invalid. There were statistically differences between preoperative score and 3 months or one year after operation. CONCLUSION: Decompression through combined posterior and anterior approaches for the treatment of pinching cervical spondyoltic myelopathy and developmental stenosis of cervical spinal canal has good efficacy, as well as a safety operation method. The operation method with one or staging decompression through combined anterior and posterior approach should be chosen according to the conditions of the patients and the operations. Anterior decompression, bone graft and titanium plate internal fixation could restore the height of vertebral body effectively and prevent fusion complication.