The oncogenic role and regulatory mechanism of PGK1 in human non-small cell lung cancer.

BACKGROUND: Phosphoglycerate kinase 1 (PGK1) is a metabolic enzyme that participates in various biological and pathological processes. Dysregulated PGK1 has been observed in numerous malignancies. However, whether and how PGK1 affects non-small cell lung cancer (NSCLC) is not yet fully elucidated. METHODS: Herein, the non-metabolic function of PGK1 in NSCLC was explored by integrating bioinformatics analyses, cellular experiments, and nude mouse xenograft models. The upstream regulators and downstream targets of PGK1 were examined using multiple techniques such as RNA sequencing, a dual-luciferase reporter assay, Co-immunoprecipitation, and Western blotting. RESULTS: We confirmed that PGK1 was upregulated in NSCLC and this upregulation was associated with poor prognosis. Further in vitro and in vivo experiments demonstrated the promoting effects of PGK1 on NSCLC cell growth and metastasis. Additionally, we discovered that PGK1 interacted with and could be O-GlcNAcylated by OGT. The inhibition of PGK1 O-GlcNAcylation through OGT silencing or mutation at the T255 O-GlcNAcylation site could weaken PGK1-mediated NSCLC cell proliferation, colony formation, migration, and invasion. We also found that a low miR-24-3p level led to an increase in OGT expression. Additionally, PGK1 exerted its oncogenic properties by augmenting ERK phosphorylation and MCM4 expression. CONCLUSIONS: PGK1 acted as a crucial mediator in controlling NSCLC progression. The miR-24-3p/OGT axis was responsible for PGK1 O-GlcNAcylation, and ERK/MCM4 were the downstream effectors of PGK1. It appears that PGK1 might be an attractive therapeutic target for the treatment of NSCLC.

GALNT2, an O-glycosylating enzyme, is a critical regulator of radioresistance of non-small cell lung cancer: evidence from an integrated multi-omics analysis.

Radioresistance is the primary reason for radiotherapy failure in non-small cell lung cancer (NSCLC) patients. Glycosylation-related alterations are critically involved in tumor radioresistance. However, the relationship between glycosylation and NSCLC radioresistance is unclear. Here, we generated radioresistant NSCLC cell models by using fractionated irradiation. The aberrant glycosylation involved in NSCLC-related radioresistance was elucidated by transcriptomic, proteomic, and glycomic analyses. We conducted in vitro and in vivo investigations for determining the biological functions of glycosylation. Additionally, its downstream pathways and upstream regulators were inferred and verified. We demonstrated that mucin-type O-glycosylation and the O-glycosylating enzyme GALNT2 were highly expressed in radioresistant NSCLC cells. GALNT2 was found to be elevated in NSCLC tissues; this elevated level showed a remarkable association with response to radiotherapy treatment as well as overall survival. Functional experiments showed that GALNT2 knockdown improved NSCLC radiosensitivity via inducing apoptosis. By using a lectin pull-down system, we revealed that mucin-type O-glycans on IGF1R were modified by GALNT2 and that IGF1R could affect the expression of apoptosis-related genes. Moreover, GALNT2 knockdown-mediated in vitro radiosensitization was enhanced by IGF1R inhibition. According to a miRNA array analysis and a luciferase reporter assay, miR-30a-5p negatively modulated GALNT2. In summary, our findings established GALNT2 as a key contributor to the radioresistance of NSCLC. Therefore, targeting GALNT2 may be a promising therapeutic strategy for NSCLC.

Financial status and travel time to driving schools as barriers to obtaining a young driver license in a state with comprehensive young driver licensing policy.

The highest lifetime risk for a motor vehicle crash is immediately after the point of licensure, with teen drivers most at risk. Comprehensive teen driver licensing policies that require completion of driver education and behind-the-wheel training along with Graduated Driver Licensing (GDL) are associated with lower young driver crash rates early in licensure. We hypothesize that lack of financial resources and travel time to driving schools reduce the likelihood for teens to complete driver training and gain a young driver's license before age 18. We utilize licensing data from the Ohio Bureau of Motor Vehicles on over 35,000 applicants between 15.5 and 25 years old collected between 2017 and 2019. This dataset of driving schools is maintained by the Ohio Department of Public Safety and is linked with Census tract-level socioeconomic data from the U.S. Census. Using logit models, we estimate the completion of driver training and license obtainment among young drivers in the Columbus, Ohio metro area. We find that young drivers in lower-income Census tracts have a lower likelihood to complete driver training and get licensed before age 18. As travel time to driving schools increases, teens in wealthier Census tracts are more likely to forgo driver training and licensure than teens in lower-income Census tracts. For jurisdictions aspiring to improve safe driving for young drivers, our findings help shape recommendations on policies to enhance access to driver training and licensure especially among teens living in lower-income Census tracts.

Clinical Effect of Uterosacral and Cardinal Ligament Fixation versus Sacrospinous Ligament Fixation of Vaginal Vault Prolapse: A Retrospective Analysis.

Objective: This study aimed at comparing sacrospinous ligament fixation (SSLF) with uterosacral and cardinal ligament fixation (USCLF) concerning complications and outcomes in patients with pelvic organ prolapse (POP). Methods: A retrospective analysis was performed on the clinical data of patients with POP stage III or above uterine prolapse treated at Wenzhou People's Hospital from January 2013 to December 2019. Patients were divided into two groups: USCLF group and SSLF group. The perioperative indicators, postoperative complications, pelvic organ prolapse quantification (POP-Q), Pelvic Floor Distress Inventory-20 (PFDI-20), and POP/Urinary Incontinence Sexual Questionnaire-12 (PISQ-12) scores of the groups were analyzed and compared. Results: (1) The operative time and intraoperative blood loss in the USCLF group were lower than those in the SSLF group, with statistical significance (p < 0.05). (2) The incidence of postoperative buttock pain in the SSLF group was 10.7% (6/56), higher than that in the USCLF group (0/56) (Fisher's exact test, p = 0.027). (3) At one year of follow-up, significant improvement in Aa, Ba, C, Ap, and Bp values was observed in both groups (p < 0.05). The values of the Aa and Ba sites in the USCLF group were lower than those in the SSLF group one year after surgery (p < 0.05). (4) The PFDI-20 and PISQ-12 scores of the groups one year after surgery were lower than those before surgery (p < 0.05). Conclusion: Uterosacral and cardinal ligament suture fixation leads to less bleeding and better postoperative quality of life than preoperative and may be better than SSLF at preventing the recurrence of anterior wall prolapse after surgery.

Comparative analysis of MVD and RHZ in the treatment of primary glossopharyngeal neuralgia: A clinical report on 61 cases.

Objective: Clinical data on 61 patients (grouped by their treatment with MVD or RHZ) with glossopharyngeal neuralgia were analyzed retrospectively. A summary analysis of the effective rate and surgical complications of MVD and RHZ in the treatment of glossopharyngeal neuralgia was performed to observe the new surgical options for GN. Method: From March 2013 to March 2020, 63 patients with GN were admitted to our hospital by the professional group of cranial nerve diseases. Two patients diagnosed with tongue and pharynx pain secondary to tongue cancer and upper esophageal cancer, respectively were excluded from the group. The remaining patients all met the diagnosis of GN, some of them were treated with MVD and others were treated with RHZ. The pain relief rate, long-term results, and complications of the patients in the two groups were well-organized and analyzed. Result: Of the 61 patients, 39 were treated with MVD and 22 were treated with RHZ. In the early-stage patients (the first 23 patients), all of them were operated on with the MVD procedure except one patient without vascular compression. In the later-stage patients, MVD was performed for evident single arterial compression according to the intraoperative situation. And for compression of arteries with greater tension or PICA + VA complex compression, RHZ was performed. It was also performed in cases where vessels with tight adhesions to the arachnoid and nerves could not be easily separated, or where it was easy to damage the perforating arteries after separating the blood vessels, causing vasospasm, which affects the blood supply to the brainstem and cerebellum. RHZ was also performed if there was no clear vascular compression. The efficiency of both groups was 100%. In the MVD group, one case recurred 4 years after the initial operation, and RHZ was performed for reoperation. Complications related to the operation included one case of swallowing and coughing in the MVD group, and three cases in the RHZ group; two cases of uvula not centering in the MVD group, and five cases in the RHZ group. There was 2 patients in RHZ group lost taste in 2/3 of the backing of the tongue, though these symptoms mostly disappeared or decreased after follow-up. One patient in the RHZ group had developed tachycardia by the time of the long-term follow-up, but whether it was related to the surgery is still uncertain. In terms of serious complications, there were two cases of postoperative bleeding in the MVD group. Based on the clinical characteristics of the patients' bleeding, it was judged that the cause of the bleeding was ischemia and was related to an intraoperative injury to the penetrating artery of the PICA artery and vasospasm. Conclusion: MVD and RHZ are effective methods for the treatment of primary glossopharyngeal neuralgia. MVD is recommended for cases where vascular compression is clear and easy to handle. However, for cases with complex vascular compression, tight vascular adhesions, difficult separation, and without clear vascular compression, RHZ could be performed. Its efficiency is equivalent to MVD, and there is no significant increase in complications such as cranial nerve disorders. There are few cranial nerve complications that seriously affect the quality of life of patients. RHZ helps to reduce the risk of ischemia and bleeding during surgery by reducing the risk of arterial spasms and injury to the penetrating arteries by separating the vessels due to separation of vessels during MVD. At the same time, it may reduce the postoperative recurrence rate.

The prediction and treatment of postpartum myofascial pelvic pain.

BACKGROUND: The clinical manifestations of myofascial pelvic pain (MFPP) are mainly acute or chronic muscle pain at one or more trigger points in the pelvic cavity or pelvic floor. OBJECTIVE: This study aims to explore the predictive value of pelvic floor myoelectric parameters with respect to MFPP and the effect of its clinical treatment. METHODS: Two hundred and one women followed up in the Wenzhou People's Hospital 6-12 weeks postpartum between July 2020 and July 2021. They were divided into an MFPP group (n= 90) and a non-MFPP group (n= 102), but 9 MFPP patients without a pelvic floor electromyography evaluation were not included. The general demographic data and pelvic floor electromyography evaluation parameters of the two groups were compared; the related factors of postpartum women suffering from MFPP were analyzed, and a nomogram model of the postpartum risk of suffering from MFPP was established. The 99 patients with postpartum MFPP were divided into a treatment group (n= 10) and a control group (n= 89). The difference in visual analog scale scores between the two groups initially and after three months of treatment was compared to evaluate the effective remission rate of postpartum MFPP after treatment. RESULTS: A significant difference was observed in the relaxation time at the rapid contraction stage (z= 4.369, p< 0.05) and the tension contraction stage (z= 135.645, p< 0.01) between the MFPP group and the non-MFPP group. The nomogram model for predicting postpartum MFPP was established with nine variables as potential predictors. The calibration chart and C index of 0.68 (95% CI: 0.65-0.71) proved that the model had a certain degree of discrimination. The clinical decision-making curve showed that the model could increase the net benefit rate of patients. The pain relief rate in the treatment group was significantly higher than that in the control group (p< 0.01). CONCLUSION: There is a significant correlation between postpartum MFPP and relaxation time at rapid contraction stage and tension contraction stage. The risk prediction nomogram model of postpartum MFPP established with nine potential predictors has a certain prediction capability, and clinical treatment can effectively relieve MFPP in postpartum patients.

The O-glycosylating enzyme GALNT2 acts as an oncogenic driver in non-small cell lung cancer.

BACKGROUND: N-Acetylgalactosaminyltransferases (GALNTs), the enzymes that initiate mucin-type O-glycosylation, are closely associated with tumor occurrence and progression. However, a comprehensive analysis of GALNTs in non-small cell lung cancer (NSCLC) is lacking. METHODS: The expression profiles and prognostic values of the GALNT family members in NSCLC were analyzed using publicly available databases. Gain- and loss-of-function experiments were applied to assess the biological function of GALNT2 in NSCLC. High-throughput sequencing and bioinformatics approaches were employed to uncover the regulatory mechanism of GALNT2. RESULTS: Among the family members of GALNTs, only GALNT2 was frequently overexpressed in NSCLC tissues and was positively correlated with poor prognosis. In vitro assays showed that GALNT2 knockdown repressed NSCLC cell proliferation, migration, and invasion, but induced apoptosis and cell cycle arrest. Correspondently, GALNT2 overexpression exerted the opposite effects. In vivo experiments demonstrated that knockdown of GALNT2 restrained tumor formation in nude mice. Mechanistic investigations revealed that GALNT2 modified the O-glycosylation of ITGA5 and affected the activation of the PI3K/Akt and MAPK/ERK pathways. Further studies showed that miR-30d was a negative regulator of GALNT2. CONCLUSIONS: These findings suggest that GALNT2 is an oncogene in NSCLC and has the potential as a target for NSCLC therapy.

Evidence from a Choice Experiment in Consumer Preference towards Infant Milk Formula (IMF) in the Context of Dairy Revitalization and COVID-19 Pandemic.

China is the largest global consumer of infant milk formula (IMF). Chinese consumer preferences towards IMF have evolved over time but have also been rocked in recent years by COVID-19 with major implications for the IMF industry, globally and within China. This study is the first to document parents' preferences toward IMF since the outbreak. We used novel methods to do so, through an online choice experiment of 804 participants that included risk perceptions and socio-demographic variables. Our study finds that Chinese parents continue to prioritize quality and safety attributes of IMF represented by functional ingredients, organic labelling and traceability information. Notably, it also finds greatly increased confidence in Chinese domestically produced IMF and an underlying preference away from expensive products. This implies that the era of 'go for foreign' and 'go for the most expensive' in IMF purchasing may be coming to an end. The shift in sentiment is driven by the longer-term revitalization of the Chinese dairy industry, accelerated by COVID-19. Understanding these trends will be of major benefit to both Chinese producers and non-Chinese exporters of IMF.

MALT1 reflects inflammatory cytokines, disease activity, and its chronological change could estimate treatment response to infliximab in Crohn's disease patients.

BACKGROUND: Mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1) mediates the immunity and inflammatory response in multiple ways to be intimately involved in the progression of autoimmune diseases. This study intended to explore the linkage of MALT1 with inflammation, disease activity, and its change with infliximab treatment response in Crohn's disease (CD) patients. METHODS: MALT1 in peripheral blood mononuclear cell samples from 72 active CD patients (at baseline, 2 weeks [W2], W6, and W12 after infliximab treatment), 20 remissive CD patients (after enrollment), and 20 healthy controls (after enrollment) were detected by RT-qPCR. RESULTS: MALT1 was highest in active CD patients, followed by remissive CD patients, and lowest in healthy controls (p < 0.001). MALT1 was positively linked with C-reactive protein (p = 0.001), erythrocyte sedimentation rate (p = 0.014), clinical disease activity index (p = 0.003), tumor necrosis factor (TNF)-α (p = 0.006), interleukin (IL)-1ß (p = 0.049), and IL-17A (p = 0.004), but not other clinical characteristics (all p > 0.05) in active CD patients. After infliximab treatment, MALT1 was decreased from baseline to W12 in active CD patients (p < 0.001), especially in responders (p < 0.001), but not in nonresponders (p = 0.053). The reduction of MALT1 at W6 (p = 0.049) and W12 (p = 0.004) was associated with a good treatment response to infliximab in active CD patients. Moreover, the response rate or MALT1 at any time point was not different between active CD patients with and without TNFi history (all p > 0.05). CONCLUSION: MALT1 reflects aggravated inflammation and disease activity. Meanwhile, the decrement of MALT1 from baseline to W12 could reflect infliximab treatment response in CD patients.

Risk of Falls in adults 45-64 years old in the United States.

Falls among the older adults (64+ years old [YO]) are considered public health issues. However, fall prevention in middle adulthood (age 45-64) has received less attention. We studied the associations between the number of falls and fall-related injuries and indicators for socio-demographics, chronic diseases, and difficulties in conducting activities in two age groups, 45-64 YO and 64+. In this secondary data analysis, we used the Behavioral Risk Factor Surveillance System (BRFSS) 2018 data. The study showed respondents in the 45-64 YO have higher average falls and fall-related injuries than those 64+ (P < .001). Variables that link to more falls and fall-related injuries in 64+ correspond to more falls and fall-related injuries in 45-64 YO. The finding indicates that the odds of falls and fall-related injuries are comparable across age groups when considering demographic characteristics. However, odds of falling in the presence of arthritis and asthma are higher for respondents in 45-64 YO than the 64+ YO. The risk of falls and fall-related injuries are not specific to older adults. Factors that matter to the number of falls and fall-related injuries in the older adults also count in the younger age group. Nurses are asked to validate available fall assessment tools for adults 45-64 years old and evaluate all clients over 45 for fall risk.

Association between high serum Nogo-B and hypertension in Chinese Han.

BACKGROUND: Cellular and animal studies have shown that endoplasmic reticulum protein B (Nogo-B) is associated with hypertension, but that association has not been fully studied in humans. Therefore, the expression levels of Nogo-B were investigated in hypertensive patients. METHODS: The plasma Nogo-B levels of 74 patients with hypertension and 67 non-hypertensive patients were measured by enzyme-linked immunosorbent assay. RESULTS: The plasma Nogo-B levels in the hypertensive group [523.43(411.41-746.79)] were higher than in the non-hypertensive group [380.29(281.57-462.13)] (P < 0.01). Pearson's correlation analysis indicated that systolic blood pressure and diastolic blood pressure were linearly and positively correlated with plasma Nogo-B levels. Multivariable logistic regression analysis was performed based on sex, age, BMI, smoking history, drinking history, and levels of TC, TG, LDL, and HDL. The results indicated that the plasma Nogo-B levels were independently associated with hypertension (OR = 1.007, 95%CI: 1.004-1.010, P < 0.01). CONCLUSIONS: The present study suggests that hypertensive participants exhibited higher plasma Nogo-B levels than those without hypertension. Plasma Nogo-B levels are independently associated with hypertension.

Cell division control 42 elevates during infliximab therapy, and its increment relates to treatment response in ulcerative colitis patients.

BACKGROUND: Cell division control 42 (CDC42) regulates multiple processes of inflammation and/or immunity in autoimmune diseases and also relates to the treatment efficacy of biologic regimens clinically. This study aimed to explore the longitudinal change in CDC42 during infliximab (IFX) treatment and its correlation with IFX response in ulcerative colitis (UC) patients. METHODS: Active UC patients (N = 48) who received IFX were recruited, and their CDC42 expressions in peripheral blood mononuclear cells (PBMCs) were detected before treatment (W0) and at 12 weeks after treatment (W12) using RT-qPCR. Also, CDC42 in PBMCs from UC patients with remission (N = 20) and health controls (HCs) (N = 20) were detected. RESULTS: CDC42 was reduced in active UC patients compared with UC patients with remission (p = 0.014) and HCs (p < 0.001). Besides, CDC42 was negatively correlated with CRP (p = 0.025), TNF-α (p = 0.024), IL-1ß (p = 0.045), IL-17A (p = 0.039), and Mayo score (p = 0.015) in active UC patients, but did not relate to ESR, disease duration, or IL-6 (all p > 0.05), while CDC42 was only negatively related to CRP in UC patients with remission (p = 0.046). Interestingly, CDC42 was increased at W12 after IFX treatment in active UC patients (p < 0.001). Specifically, CDC42 was elevated during treatment in active UC patients with IFX response (p < 0.001), but did not obviously change in those without IFX response (p = 0.061). Furthermore, CDC42 at W12 was higher in active UC patients with IFX response compared with those without IFX response (p = 0.049). CONCLUSION: Cell division control 42 serves as a potential biomarker for monitoring disease progression and IFX response in UC patients.

Impact of TNC on travel behavior and mode choice: a comparative analysis of Boston and Philadelphia.

We compare responses from an online survey among 700 customers of transportation network companies (TNC) in Boston and Philadelphia to investigate TNC's impact on vehicle ownership, trip making, and mode choice. We first use a qualitative comparative analysis to examine changes in respondents' travel behavior and vehicle ownership after adopting TNC. We then use a random parameter logit regression analysis to investigate customers' preferences between transit and TNC based on a choice experiment. We find that in both cities, TNC allows customers, including those who currently do not own a car, to either delay purchasing a car or forgo a car altogether. TNC enables customers across income levels to take trips that they otherwise would not have taken. Meanwhile, TNC substitutes for more than complementing transit. The random parameter logit analysis indicates that when choosing between TNC and transit, individuals in both cities consider waiting time and overall travel time for transit to be more burdensome than those for TNC. Bostonians perceive the time spent walking to and from transit to be less burdensome, and the time spent traveling in vehicle to be more burdensome than do Philadelphians. Differences in built environment, mode share within transit systems, and income likely contribute to respondents' different values of time between the two cities. Our paper is the first to compare individual trade-off between transit and TNC in two cities with different urban settings and transit services. The findings have implications on transit service planning, station area improvements, parking regulations, and traffic management. Supplementary Information: The online version contains supplementary material available at 10.1007/s11116-021-10220-5.

A novel mechanism for C1GALT1 in the regulation of gastric cancer progression.

BACKGROUND: Gastric cancer (GC) is a highly aggressive and lethal disease around the world. High expression of core 1 ß 1, 3-galactosyltransferase 1 (C1GALT1), the primary enzyme responsible for protein O-glycosylation, plays a critical role in gastric carcinogenesis. However, proteins that can be O-glycosylated by C1GALT1 in GC have not been completely elucidated. Also, the mechanism leading to its upregulation in GC is currently unknown. RESULTS: Using public databases and our patient samples, we confirmed that C1GALT1 expression was upregulated at both the mRNA and protein levels in GC tissues. Elevated expression of C1GALT1 protein was closely associated with advanced TNM stage, lymph node metastasis, tumor recurrence, and poor overall survival. With gain- and loss-of-function approaches, we demonstrated that C1GALT1 promoted GC cell proliferation, migration, and invasion. By employing lectin pull-down assay and mass spectrometry, integrin α5 was identified as a new downstream target of C1GALT1 in GC. C1GALT1 was able to modify O-linked glycosylation on integrin α5 and thereby modulate the activation of the PI3K/AKT pathway. Functional experiments indicated that integrin α5 inhibition could reverse C1GALT1-mediated tumor growth and metastasis both in vitro and in vivo. Moreover, transcription factor SP1 was found to bind to the C1GALT1 promoter region and activated its expression. Further investigation proved that miR-152 negatively regulated C1GALT1 expression by directly binding to its 3' -UTR. CONCLUSIONS: Our findings uncover a novel mechanism for C1GALT1 in the regulation of GC progression. Thus, C1GALT1 may serve as a promising target for the diagnosis and treatment of GC.

C1GALT1, Negatively Regulated by miR-181d-5p, Promotes Tumor Progression via Upregulating RAC1 in Lung Adenocarcinoma.

Glycosyltransferases are frequently dysregulated in lung cancer. Core 1 ß 1, 3-galactosyltransferase 1 (C1GALT1), an enzyme highly expressed in various cancers, is correlated with tumor initiation and development. However, the role of C1GALT1 in lung cancer remains poorly understood. In this study, through bioinformatic analysis and clinical validation, we first discovered that C1GALT1 expression was upregulated in lung adenocarcinoma (LUAD) tissues and was closely related to poor prognosis in patients with LUAD. Gain- and loss-of-function experiments showed that C1GALT1 promoted LUAD cell proliferation, migration, and invasion in vitro, as well as tumor formation in vivo. Further investigation demonstrated that RAC1 expression was positively regulated by C1GALT1 in LUAD, whereas silencing Rac1 could reverse C1GALT1-induced tumor growth and metastasis. Moreover, miR-181d-5p was identified as a negative regulator for C1GALT1 in LUAD. As expected, the inhibitory effects of miR-181d-5p on LUAD cell proliferation, migration, and invasion were counteracted by restoration of C1GALT1. In summary, our results highlight the importance of the miR-181d-5p/C1GALT1/RAC1 regulatory axis during LUAD progression. Thus, C1GALT1 may serve as a potential therapeutic target for LUAD.

The efficacy of manipulation as a treatment for myofascial pelvic pain.

OBJECTIVE: To evaluate the effect of myofascial manipulation by observing the changes in pelvic floor myofascial scores and electromyography (EMG) data before and after treatment. METHODS: A total of 106 patients with myofascial pelvic pain (MFPP) were enrolled in a treatment group, and 50 healthy women were enrolled in a control group. The changes in the pelvic floor EMG data in the two groups were monitored by using Myo Trac before and after treatment. Pelvic trigger points and their distribution in the MFPP patients were examined using a finger pressure test. The visual analogue scale was used to assess the severity of pain in both groups. After one course of manipulation (twice per week for a total of 10 times), the effectiveness of the manipulation was analyzed by comparing the changes in pain scores before and after treatment. RESULTS: The main symptoms of MFPP in the study sample consisted of lower abdominal pain, lumbosacral pain, or mixed pain, which together accounted for 67% of all symptoms. Patients often had multiple trigger points, covering 47.17% of the body. The differences between the treatment group and control group in the changes in pelvic floor muscle strength, number of pain points, pain scores, resting EMG of pelvic floor muscles, and relaxation time after muscle contraction were all statistically significant (P < 0.05). The differences between the pre-treatment and post-treatment groups in the changes in pelvic floor muscle strength, number of pain points, pain scores, resting EMG of pelvic floor muscles, and relaxation time after muscle contraction were all statistically significant (P < 0.05) CONCLUSION: Manipulation is an effective treatment for MFPP and is worthy of further clinical promotion.

Identification of LCA-binding Glycans as a Novel Biomarker for Esophageal Cancer Metastasis using a Lectin Array-based Strategy.

Esophageal cancer (EC) is a unique and heterogeneous disease diagnosed mostly at advanced stages. Altered glycans presented on cell surfaces are involved in the occurrence and development of malignancy. However, the effects of glycans on EC progression are largely unexplored. Here, a lectin array was utilized to detect the glycan profiling of the normal esophageal mucosal epithelial cell line and two EC cell lines. The binding of Lens culinaris lectin (LCA) to EC cells was found to be stronger than that of the normal cells. Lectin immunohistochemical staining revealed that LCA-binding glycans were markedly elevated in EC tissues compared to adjacent non-cancerous tissues. LCA staining was significantly associated with lymph node metastasis, depth of invasion, TNM stage and poor overall survival of EC patients. Added LCA to block LCA recognized glycans could inhibit the migration and invasion of EC cells. Further analysis revealed that blocking the biosynthesis of LCA-binding glycans by tunicamycin attenuated cellular migratory and invasive abilities. Additionally, a membrane glycoprotein CD147 was recognized as a binder of LCA. There was a positive correlation between LCA-binding glycans and CD147 expression in clinical samples. Interestingly, CD147 inhibition also reduced cell migration and invasion. These findings indicated that LCA-binding glycans may function as a novel indicator to predict metastasis for patients with EC.

A lectin-based glycomic approach identifies FUT8 as a driver of radioresistance in oesophageal squamous cell carcinoma.

PURPOSE: Radio-resistance is recognized as a main factor in the failure of radiotherapy in oesophageal squamous cell carcinoma (ESCC). Aberrant cell surface glycosylation has been reported to correlate with radio-resistance in different kinds of tumours. However, glycomic alterations and the corresponding enzymes associated with ESCC radio-resistance have not yet been defined. METHODS: Two radioresistant cell lines, EC109R and TE-1R, were established from parental ESCC cell lines EC109 and TE-1 by fractionated irradiation. A lectin microarray was used to screen for altered glycan patterns. RNA-sequencing (RNA-seq) was employed to identify differentially expressed glycosyltransferases. Cell Counting Kit-8, colony formation and flow cytometry assays were used to measure cell viability and radiosensitivity. Expression of glycosyltransferase in ESCC tissues was assessed by immunohistochemistry. In vivo radiosensitivity was analysed using a nude mouse xenograft model. Downstream effectors of the enzyme were verified using a lectin-based pull-down assay combined with mass spectrometry. RESULTS: We found that EC109R and TE-1R cells were more resistant to irradiation than the parental EC109 and TE-1 cells. Using lectin microarrays combined with RNA sequencing, we found that α1, 6-fucosyltransferase (FUT8) was overexpressed in the radioresistant ESCC cell lines. Both gain- and loss-of-function studies confirmed that FUT8 regulates the sensitivity of ESCC cells to irradiation. Importantly, we found that high FUT8 expression was positively linked to radio-resistance and a poor prognosis in ESCC patients who received radiation therapy. Moreover, FUT8 inhibition suppressed the growth and formation of xenograft tumours in nude mice after irradiation. Using a lectin-based pull-down assay and mass spectrometry, we found that CD147 could be glycosylated by FUT8. As expected, inhibition of CD147 partly reversed FUT8-induced radio-resistance in ESCC cells. CONCLUSIONS: Our results indicate that FUT8 functions as a driver of radio-resistance in ESCC by targeting CD147. Therefore, FUT8 may serve as a marker for predicting the response to radiation therapy in patients with ESCC.

RRFT1 (Redox Responsive Transcription Factor 1) is involved in extracellular ATP-regulated gene expression in Arabidopsis thaliana seedlings.

As an apoplast signal molecule, extracellular ATP (eATP) is involved in the growth regulation of Arabidopsis thaliana seedlings. Recently, RRFT1 was revealed to be involved in eATP- regulated seedling growth. To further verify the role of RRTF1 in seedlings' eATP response, expression of 20 eATP-responsive genes in wild type (Col-0) and RRTF1 null mutant (rrtf1-1) seedlings were investigated by using realtime quantitative PCR. After 0.5 mM ATP stimulation, the response of these genes' expression in rrtf1-1 seedlings was significantly different from that in Col-0 seedlings. Proteins which are encoded by these genes include transcription factors, plasma membrane receptors like kinases, ion influx/efflux transporters and hormone signaling components. The results indicated that RRTF1 may be involved in eATP regulated physiological responses via regulating the expression of some functional genes.