Systemic immune-inflammation index in predicting non-curative resection of endoscopic submucosal dissection in patients with early gastric cancer.

BACKGROUND AND PURPOSE: Although endoscopic submucosal dissection (ESD) is considered standard treatment for early gastric cancer (EGC), patients with non-curative resection (NCR) of ESD may still require gastrectomy. The systemic immune-inflammation index (SII) showed great potential in predicting the prognosis of gastric cancer patients. This study aims to investigate the predictive validity of SII of NCR in EGC patients. METHODS: We reviewed data from EGC patients who underwent ESD in the past. The relationship between SII and clinicopathologic features was investigated. We used Receiver operating characteristic curves to compare the predictive values of NCR between SII and other inflammation indices. Binary logistic analysis was used to identify independent risk factors for NCR. These factors were then used to construct a predictive nomogram. RESULTS: SII was associated with larger tumor size, male gender, older age, submucosal invasion, and a greater risk of NCR. SII showed better predictivity of NCR than platelet-to-lymphocyte ratio (PLR) and neutrophil-to-lymphocyte ratio (NLR). SII [odds ratio (OR) = 1.003, P = 0.001], NLR (OR = 1.520, P = 0.029), PLR (OR = 1.009, P = 0.010), upper stomach tumors (OR = 16.393, P < 0.001), poorly differentiated type (OR = 29.754, P < 0.001), ulceration (OR = 4.814, P = 0.001), and submucosal invasion (OR = 48.91, P < 0.001) were independent risk factors for NCR. The nomogram model based on these factors exhibited superior concordance and accuracy. CONCLUSION: SII could be considered a simple and effective predictor of NCR of ESD in EGC patients.

Physical activity levels of children and adolescents with intellectual disabilities in Northern China.

BACKGROUND: Limited research has looked at the physical activity (PA) among Chinese children and adolescents with intellectual disabilities. METHODS: The Children's Leisure Activities Study Survey-Chinese version was applied in evaluating PA levels among children and adolescents with intellectual disabilities. RESULTS: The data indicated that only 29.5% of children and adolescents with intellectual disabilities met the PA recommendation. Boys engaged in significantly more MVPA and had higher energy expenditure than girls during weekdays and the whole week. The general trend in PA levels demonstrated a downtrend with the growth of age of children and adolescents with intellectual disabilities. The students aged 6-12 years old performed more MVPA and had greater energy expenditure compared with those 13-15 and 16-18 years old. CONCLUSION: The level of PA among Chinese children and youths with intellectual disabilities is remarkably low, and there is a large gap compared with the PA recommendation.

Effect of Interactive Dynamic Scalp Acupuncture on Post-Stroke Cognitive Function, Depression, and Anxiety: A Multicenter, Randomized, Controlled Trial.

OBJECTIVE: To compare the clinical effects of interactive dynamic scalp acupuncture (IDSA), simple combination therapy (SCT), and traditional scalp acupuncture (TSA) on cognitive function, depression and anxiety in patients with post-stroke cognitive impairment. METHODS: A total of 660 patients with post-stroke cognitive impairment who were admitted to 3 hospitals in Shenzhen City between May 2017 and May 2020 were recruited and randomly assigned to the IDSA (218 cases), SCT (222 cases) and TSA groups (220 cases) according to a random number table. All the patients received conventional drug therapy for cerebral stroke and exercise rehabilitation training. Scalp acupuncture and computer-based cognitive training (CBCT) were performed simultaneously in the IDSA group, but separately in the morning and in the afternoon in the SCT group. The patients in the TSA group underwent scalp acupuncture only. The course of treatment was 8 weeks. Before treatment (M0), 1 (M1) and 2 months (M2) after treatment, as well as follow-up at 1 (M3) and 2 months (M4), the cognitive function of patients was assessed using the Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment Scale (MoCA) Scales; depression, anxiety, sleep quality, and self-care ability of patients were assessed using Hamilton Depression Rating Scale (HAMD), Hamilton Anxiety Rating Scale (HAMA), Pittsburgh Sleep Quality Index (PSQI), and Modified Barthel Index (MBI), respectively. During this trial, all adverse events (AEs) were accurately recorded. RESULTS: There were no significant differences in the MMSE, MoCA, HAMD, HAMA, PSQI, and MBI scores among the 3 groups at M0 (all P>0.05). In the IDSA group, the MMSE, MoCA and MBI scores from M2 to M4 were significantly higher than those in the SCT and TSA groups, while the HAMD, HAMA and PSQI scores were significantly reduced (all P<0.01). The changes of all above scores (M2-M0, M4-M0) were significantly superior to those in the SCT and TSA groups (all P<0.01, except M4-M0 of HAMD). At M2, the severity of MMSE, HAMD, HAMA, PSQI and MBI in the IDSA group was significantly lower than that in the SCT and TSA groups (all P<0.01). There was no serious AE during this trial. CONCLUSIONS: IDSA can not only significantly improve cognitive function, but also reduce depression, anxiety, which finally improves the patient's self-care ability. The effect of IDSA was significantly better than SCT and TSA. (Trial registration No. ChiCTR1900027206).

Effects of Interactive Dynamic Scalp Acupuncture on Motor Function and Gait of Lower Limbs after Stroke: A Multicenter, Randomized, Controlled Clinical Trial.

OBJECTIVE: To evaluate the effects of interactive dynamic scalp acupuncture (IDSA), simple combination therapy (SCT), and traditional scalp acupuncture (TSA) on motor function and gait of the lower limbs in post-stroke hemiplegia patients. METHODS: A total of 231 patients with post-stroke hemiplegia was randomly divided into IDSA (78 cases), SCT (78 cases), and TSA (75 cases) groups by a random number table. Scalp acupuncture (SA) and lower-limb robot training (LLRT) were both performed in the IDSA and SCT groups. The patients in the TSA group underwent SA and did not receive LLRT. The treatment was administered once daily and 6 times weekly for 8 continuous weeks, each session lasted for 30 min. The primary outcome measures included Fugl-Meyer assessment of the lower extremity (FMA-LE), berg balance scale (BBS), modified barthel index (MBI), and 6-min walking test (6MWT). The secondary outcome measures included stride frequency (SF), stride length (SL), stride width (SW), affected side foot angle (ASFA), passive range of motion (PROM) of the affected hip (PROM-H), knee (PROM-K) and ankle (PROM-A) joints. The patients were evaluated before treatment, at 1- and 2-month treatment, and 1-, and 2-month follow-up visits, respectively. Adverse events during 2-month treatment were observed. RESULTS: Nineteen patients withdrew from the trial, with 8 in the IDSA and 5 in the SCT groups, 6 in the TSA group. The FMA-LE, BBS, 6MWT and MBI scores in the IDSA group were significantly increased after 8-week treatment and 2 follow-up visits compared with the SCT and TSA groups (P<0.05 or P<0.01). Compared with pre-treatment, the grade distribution of BBS and MBI scores in the 3 groups were significantly improved at 1, 2-month treatment and 2 follow-up visits (P<0.05 or P<0.01). The SF, PROM-H, PROM-K and PROM-A in the IDSA group was significantly increased compared with the SCT and TSA groups after 8-week of treatment (P<0.05 or P<0.01). Compared with the SCT group, ASFA of the IDSA group was significantly reduced after 8-week of treatment (P<0.05). SF, SL, PROM-K and PROM-A were significantly increased at the 2nd follow-up visit whereas the ASFA was significantly reduced in the IDSA group compared with the SCT groups at 1st follow-up visit (P<0.05 or P<0.01). The SF was significantly increased in the SCT group compared with the TSA group after 8-week treatment (P<0.05). Compared with the TSA group, PROM-K, PROM-A were significantly increased at the 2nd follow-up visit (P<0.05). CONCLUSIONS: The effects of IDSA on lower-limb motor function and walking ability of post-stroke patients were superior to SCT and TSA. The SCT was comparable to TSA treatment, and appeared to be superior in improving the motion range of the lower extremities. (Registration No. ChiCTR1900027206).

[Lycium barbarum polysaccharide reduces testicular spermatogenic injury in Immp2l－/－mice through GPX4 and AIF pathways].

OBJECTIVE: To investigate the protective effect of Lycium barbarum polysaccharide (LBP) against testicular spermatogenic injury in mice with oxidative stress (OS) and its mechanism. METHODS: A unique OS model was made in 1.5-month-old mice with mitochondrial inner membrane-like peptide-2 mutation (Immp2l－/－), which were fed with water (the negative control group) or LBP in water at the concentration of 20 mg/kg (the LBP intervention group), and wild-type Immp2l＋/＋ mice used as normal controls and fed with water only. Then all the mice were sacrificed at 13 months old and the testis tissue harvested for observation of pathological changes by HE staining, measurement of routine semen parameters, and detection of the apoptosis of spermatogenic cells by TUNEL and the expression levels of glutathione peroxidase 4 (GPX4) and apoptosis-inducing factor (AIF) by immunohistochemistry and Western blot. RESULTS: Thinned testicular cortex was observed in the negative controls, with evident vacuolar degeneration and reduced numbers of germ cells and elongated spermatids in the lumen of the seminiferous tubules, but all these pathological changes were improved and the germ cells at different levels orderly arranged in the LBP intervention group. Compared with the normal controls, the mice in the negative control group showed dramatically reduced sperm count (ï¼»72.89 ± 8.28ï¼½ vs ï¼»20.78 ± 1.45ï¼½ ×106, P<0.01) and the percentages of progressively motile sperm (PMS) (ï¼»58.62 ± 6.15ï¼½% vs ï¼»18.37 ± 2.67ï¼½%, P<0.01) and morphologically normal sperm (MNS) (ï¼»65.81 ± 7.69ï¼½% vs ï¼»20.33 ± 3.17ï¼½%, P<0.01) and increased apoptosis of spermatogenic cells (ï¼»1.45 ± 0.43ï¼½% vs ï¼»7.14 ± 0.78ï¼½%, P<0.01). LBP intervention, however, significantly increased the sperm count (ï¼»45.25 ± 3.39ï¼½ ×106, P<0.05), PMS (ï¼»36.34 ± 4.56ï¼½%, P<0.05) and MNS (ï¼»38.72 ± 3.63ï¼½%, P<0.05) and decreased the apoptosis of spermatogenic cells (ï¼»2.28 ± 0.07ï¼½%, P<0.01). The mice in the LBP intervention group, in comparison with the negative controls, exhibited remarkably up-regulated expression of GPX4 (2.75 ± 0.48 vs 1.43 ± 0.17, P<0.05) and down-regulated expression of AIF (2.43 ± 0.15 vs 1.35 ± 0.51, P<0.05). CONCLUSIONS: Lycium barbarum polysaccharide at 20 mg/kg can reduce testicular spermatogenic injury in Immp2l－/－ mice with oxidative stress through GPX4 and AIF pathways.

Efficacy and safety of low-dose everolimus combined with endocrine drugs for patients with hormone receptor-positive, human epidermal growth factor receptor 2-negative metastatic breast cancer.

BACKGROUND: To analyze the efficacy and safety of everolimus 5 mg/day in combination with endocrine drugs in the treatment of hormone receptor-positive, human epidermal growth factor receptor 2 (HER2)-negative metastatic breast cancer using real-world clinical data. METHODS: Clinical data of hormone receptor (HR)-positive and HER2-negative patients with advanced breast cancer treated with everolimus combined with endocrine drugs in our center between August 2012 and May 2017 were retrospectively analyzed. Curative effect and adverse reactions were evaluated. RESULTS: A total of 110 patients were enrolled in this study, and 87.3% received salvage chemotherapy. The median number of salvage treatment lines was 5 (range: 1-19). The median follow-up duration was 12 months (range: 1-56.3 months), the overall response rate (ORR) was 6.4%, the clinical benefit rate (CBR) was 31.8%, the median progression-free survival (mPFS) was 4.0 months (95% CI: 2.9-5.1 months), and the median overall survival (OS) was 17 months (95% CI: 12.1-21.9 months). The mPFS for patients who received ≤2 treatment line was 11.8 months (95% CI: 4.3-19.3 months). Univariate and multivariate analyses suggested that absence of liver metastases, secondary endocrine resistance, and number of metastasis sites <3 were the main factors influencing the benefit of everolimus combined with endocrine therapy. The most common adverse events of grade 3 were: stomatitis (5.5%), non-infectious pneumonia (1.8%), and erythra (1.8%). No grade 4 adverse reactions were observed. CONCLUSIONS: Our results showed that everolimus (5 mg/day) combined with endocrine therapy was effective and relatively safe for patients with hormone receptor-positive, HER2-negative metastatic breast cancer.

Corrigendum: Neutrophil-to-Lymphocyte and Platelet-to-Lymphocyte Ratios as Predictors of Outcomes in Patients With Unresectable Hepatocellular Carcinoma Undergoing Transarterial Chemoembolization Plus Sorafenib.

[This corrects the article DOI: 10.3389/fmolb.2021.624366.].

Neutrophil-to-Lymphocyte and Platelet-to-Lymphocyte Ratios as Predictors of Outcomes in Patients With Unresectable Hepatocellular Carcinoma Undergoing Transarterial Chemoembolization Plus Sorafenib.

Objectives: To investigate the predictive value of inflammatory biomarkers in patients with unresectable hepatocellular carcinoma (HCC) for outcomes following the combination treatment of transarterial chemoembolization (TACE) plus sorafenib. Materials and Methods: A total of 314 (270 male and 44 female) treatment-naïve patients with unresectable HCC treated by TACE plus sorafenib between January 2011 and December 2018 were enrolled in the retrospective study. The primary outcome was overall survival (OS). The secondary outcome was progression-free survival (PFS). Neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) were obtained within 3-7 days before the initial TACE and the median value of the NLR and PLR was considered as the cut-off value. Results: The median value of NLR and PLR was 2.42 and 100, respectively. The median OS and PFS of the entire cohort were 18.7 months (95% CI: 16.8-20.6) and 9.1 months (95% CI: 8.5-9.8), respectively. The low NLR and PLR group showed improved OS and PFS compared with the high NLR and PLR group [21.8 months (95% CI: 15.2-28.5) vs. 15.4 months (95% CI: 12.4-18.3), p < 0.0001; 21.6 months (95% CI: 15.8-27.5) vs. 14.9 months (95% CI: 11.9-17.8), p = 0.00027, respectively]. In addition, the low NLR and PLR group also provided a longer PFS than the high NLR and PLR group [10.4 months (95% CI: 8.9-12.0) vs. 8.1 months (95% CI: 7.1-9.2), p = 0.00022; 10.3 months (95% CI: 8.6-11.9) vs. 8.2 months (95% CI: 7.2-9.2), p < 0.0001, respectively]. High NLR and PLR at baseline were predictive factors of poor OS (p = 0.02 and p = 0.004) and PFS (p = 0.045 and p = 0.005). Conclusion: This study showed the prognostic value of quantitative inflammatory biomarkers in correlation with OS and PFS in unresectable HCC patients undergoing TACE plus sorafenib treatment.

[Scalp acupuncture combined with lower-limb intelligent feedback training for lower-limb motor dysfunction after stroke: a randomized controlled trial].

OBJECTIVE: To compare the efficacy of scalp acupuncture combined with lower-limb intelligent feedback training and lower-limb intelligent feedback training alone for lower-limb motor dysfunction after stroke. METHODS: A total of 154 patients with lower-limb motor dysfunction after stroke were randomly divided into an observation group (76 cases, 6 cases dropped off) and a control group (78 cases, 8 cases dropped off). The patients in both groups were treated with conventional medication and exercise rehabilitation training. In addition, the patients in the observation group were treated with scalp acupuncture combined with lower-limb intelligent feedback training. The scalp acupuncture was given at upper 1/5 of the anterior oblique line of parietal temporal area and upper 1/5 of the posterior oblique line of parietal temporal area. The patients in the control group were treated with lower-limb intelligent feedback training alone. All the treatment was given once a day, 6 days a week, totaling for 8 weeks. The affected-side lower-limb Brunnstrom stage and modified Ashworth scale (MAS) grade, 6-minute walk test (6MWT), Berg balance scale (BBS) score and modified Barthel index (MBI) score were evaluated before and after treatment in the two groups. The plantar pressure was measured by gait function evaluation system. RESULTS: Compared before treatment, the Brunnstrom stage in the two groups was improved after treatment (P<0.01); the MAS grade in the observation group was improved after treatment (P<0.01); the Brunnstrom stage and MAS grade in the observation group were superior to those in the control group (P<0.01, P<0.05). After treatment, the 6MWT, BBS and MBI scores in the two groups were increased (P<0.05), and those in the observation group were higher than those in the control group (P<0.05). After treatment, the touchdown area of health-side hind foot, affected-side front-hind foot and bilateral full foot in the observation group was increased (P<0.05), and the touchdown area of affected-side front-hind foot and full foot in the observation group was larger than that in the control group (P<0.05). The weight-bearing ratio of health-side forefoot and full foot in the observation group was decreased after treatment (P<0.05), and the weight-bearing ratio of affected-side forefoot, hind foot and full foot was increased after treatment (P<0.05). The weight-bearing ratio of health-side forefoot and full foot in the observation group was lower than that in the control group (P<0.05), and the weight-bearing ratio of health-side hind foot, affected-side forefoot and affected-side full foot in the observation group was higher than that in the control group (P<0.05). CONCLUSION: The scalp acupuncture combined with lower-limb intelligent feedback training could reduce the muscle tension of lower limbs, promote the separation movement mode of lower limbs, improve the plantar pressure distribution, and improve the balance ability and walking ability in stroke patients, and the curative effect is better than lower-limb intelligent feedback training alone.

[Interactive scalp acupuncture for cognitive dysfunction after stroke: a randomized controlled trial].

OBJECTIVE: To compare the efficacy of interactive scalp acupuncture, scalp acupuncture alone and scalp acupuncture plus cognitive training for cognitive dysfunction after stroke. METHODS: A total of 660 patients with cognitive dysfunction after stroke were randomly divided into an interactive scalp acupuncture group (218 cases, 18 cases dropped off), a scalp acupuncture group (220 cases, 20 cases dropped off) and a scalp acupuncture plus cognitive training group (222 cases, 22 cases dropped off). All the patients were treated with routine medication and exercise rehabilitation training. The interactive scalp acupuncture group was treated with scalp acupuncture on the parietal midline, and contralateral anterior parietal temporal oblique line and posterior parietal temporal oblique line at the same time of cognitive training; the scalp acupuncture group was treated with scalp acupuncture alone, and the scalp acupuncture plus cognitive training group was treated with scalp acupuncture and cognitive training in the morning and afternoon respectively. All the treatments were given once a day, 6 times a week for 8 weeks. Montreal cognitive assessment (MoCA) scale score was used to evaluate the cognitive function before treatment, 4 weeks and 8 weeks into treatment. RESULTS: Compared before treatment, the total score of MoCA was increased after 4-week treatment and 8-week treatment in the three groups (P<0.01), and the score in the interactive scalp acupuncture group was higher than that in the scalp acupuncture group and the scalp acupuncture plus cognitive training group (P<0.05, P<0.01). Compared before treatment, each item score of MoCA was increased after 8-week treatment in the three groups (P<0.01), and the score in the interactive scalp acupuncture group was higher than that in the scalp acupuncture group and the scalp acupuncture plus cognitive training group (P<0.01). Except for the attention, the remaining items scores of MoCA in the scalp acupuncture plus cognitive training group were higher than those in the scalp acupuncture group (P<0.01). CONCLUSION: The interactive scalp acupuncture could significantly improve the cognitive function in patients with cognitive dysfunction after stroke, and the efficacy is superior to scalp acupuncture alone and scalp acupuncture plus cognitive training.

[Characteristics and Removal Mechanism of an Electro-Hybrid Ozonation-Coagulation System in the Treatment of Organic Matters].

To improve the removal efficiency of dissolved organic matter in wastewater treatment plant (WWTP) effluent, electro-hybrid ozonation-coagulation (E-HOC) is proposed and the treatment characteristics and removal mechanism for WWTP effluent and ibuprofen (IBP) are investigated. The E-HOC process has a better removal effect on dissolved organic matter in WWTP effluent, achieving 46.4%, 20.0%, 19.4%, 36.1%, and 49.7% higher removal than EC, ozonation, pre-ozonation-EC, electrocoagulation-ozonation, and chemical coagulation, respectively. To determine the mechanism of the E-HOC process, quenching experiments and electron paramagnetic resonance (EPR) were conducted, which confirmed that metal coagulants can be used as a catalyst to effectively increase the generation of the hydroxyl radical (·OH). Synergistic effects between ozone and the coagulants (SOC) were also found to be involved. Fourier-transform infrared spectroscopy (FT-IR) illustrated that the surface hydroxyl groups of the coagulant (hydrolyzed species produced by Al anode electrolysis) were the active sites for the generation of·OH in the SOC reaction. Based on a kinetics analysis of organic matter removal in the E-HOC system, SOC effects and ozonation played dominant roles in the E-HOC process. Additionally, the SOC created a new pathway for·OH formation.

Systematic Discovery and Pathway Analyses of Metabolic Disturbance in COVID-19.

Background: The ongoing global coronavirus disease 2019 (COVID-19) pandemic is posing a serious public health threat to nations worldwide. Understanding the pathogenesis of the disease and host immune responses will facilitate the discovery of therapeutic targets and better management of infected patients. Metabolomics technology can provide an unbiased tool to explore metabolic perturbation. Methods: Twenty-six healthy controls and 50 COVID-19 patients with mild, moderate, and severe symptoms in the Fifth Medical Center of PLA General Hospital from January 22 to February 16, 2020 were recruited into the study. Fasting blood samples were collected and subject to metabolomics analysis by liquid chromatography-mass spectrometry. Metabolite abundance was measured by peak area and was log-transformed before statistical analysis. The principal component analysis, different expression analysis, and metabolic pathway analysis were performed using R package. Co-regulated metabolites and their associations with clinical indices were identified by the weighted correlation network analysis and Spearman correlation coefficients. The potential metabolite biomarkers were analyzed using a random forest model. Results: We uncovered over 100 metabolites that were associated with COVID-19 disease and many of them correlated with disease severity. Sets of highly correlated metabolites were identified and their correlations with clinical indices were presented. Further analyses linked the differential metabolites with biochemical reactions, metabolic pathways, and biomedical MeSH terms, offering contextual insights into disease pathogenesis and host responses. Finally, a panel of metabolites was discovered to be able to discriminate COVID-19 patients from healthy controls, and also another list for mild against more severe cases. Our findings showed that in COVID-19 patients, citrate cycle, sphingosine 1-phosphate in sphingolipid metabolism, and steroid hormone biosynthesis were downregulated, while purine metabolism and tryptophan metabolism were disturbed. Conclusion: This study discovered key metabolites as well as their related biological and medical concepts pertaining to COVID-19 pathogenesis and host immune response, which will facilitate the selection of potential biomarkers for prognosis and discovery of therapeutic targets.

[Metagenomic Insights into Salinity Build-up in Microbial Communities and Metabolism of Hydrolytic Bioreactor Treating High-color PDWW].

In this study, to solve the problem of salinity enrichment in industrial wastewater recycling, a hydrolytic bioreactor was continuously operated to treat high-color printing and dyeing wastewater (PDWW) with salinity build-up. Nearly complete color removal was achieved even with salinity build-ups from 0.5 to 4 g·L-1 in the influent. Pyrosequencing of 16S rRNA genes showed that the salinity build-up results in the decrease of microbial species from 882 to 631; however, the biodiversity of the bacterial community remains stable. Metagenomic analysis indicated that salinity build-up caused no obvious effect on the overall function of the bacterial community, but altered the abundance of specific decoloring genes. Proteobacteria dominated in the bioreactor, and Methanothrix and Geobacter were the dominant genera under low salinity conditions. Proteobacteria increased in abundance with salinity build-up. Desulfovibrio and Desulfococcus were the two predominant genera in the bioreactor fed with sodium sulphate salinity build-up, demonstrating opposite responses to the sodium stress. PICRUSt functional analysis showed that the relative abundance of the decolorizing enzymes SOD1 and SOD2 decreased significantly, but the relative abundance of CAT and TYR increased, ensuring the stability of the decolorizing function of the hydrolysis biological system. From the perspective of the functional genes of hydrolysis decolorization, this study explored the effect of salinity build-up on the microbial community and function of hydrolysis, providing a theoretical basis for the study of decolorization and organic matter removal mechanism of PDWW under the condition of salinity build-up.

CaMKKß regulates proliferation, apoptosis, and glycolysis of hepatocellular carcinoma via PI3K/AKT pathway.

BACKGROUND: Hepatocellular carcinoma (HCC) is the most common malignancy of liver cancer. Calcium ions/calmodulins stimulate protein kinase kinases ß (CaMKKß) is a multifunctional protein kinase that is overexpressed in many types of cancer. This study aims to investigate the effect of CaMKKß interference on HCC in HepG2 cells and transplanted tumor mice. METHODS: CaMKKß gene was knocked out in HepG2 cells as an experimental group, empty vector lentivirus as a negative control (NC) group, and untreated HepG2 cells as a control group. Cell proliferation, cell cycle, apoptosis, invasion, and glycolysis potential assays were conducted, respectively. In addition, the expression of PI3K, p-PI3K, AKT, and p-AKT was quantified by Western blot. Finally, the effect of CaMKKß in vivo was investigated using a xenograft model. RESULTS: CaMKKß knockdown significantly suppressed HepG2 cell proliferation, cell cycle, invasion, EMT, and glycolysis, promoted cell apoptosis, and reduced the expression of hexokinase 2 (HK2), pyruvate kinase M (PKM2), and lactate dehydrogenase A (LDHA), p-PI3K, and p-AKT. Post the addition of AKT highly expression plasmid, glucose uptake, lactic acid production, and cell proliferation decreased, accompanied by an increase in apoptosis, which were substantially reversed. Notably, xenograft model experiments in vivo also confirmed that CaMKKß knockdown inhibited HCC growth. CONCLUSIONS: CaMKKß knockdown inhibited cell proliferation, invasion, and glycolysis through the PI3K/AKT pathway, heightened apoptosis, thus promoting the development of HCC. This might be a potential target for the diagnosis and treatment of HCC.

Characterization of exosome-like vesicles derived from Taenia pisiformis cysticercus and their immunoregulatory role on macrophages.

BACKGROUND: Taenia pisiformis is one of the most common intestinal parasites in canines, and leads to serious economic losses in the rabbit breeding industry. Exosome-like vesicles from parasites play crucial roles in host-parasite interactions by transferring cargo from parasites to host cells and by modulating host immunological response through inducing production of host-derived cytokines. Nevertheless, the mechanism by which exosome-like vesicles from T. pisiformis cysticercus regulate the macrophage immune response remains unknown. METHODS: Using ultracentrifugation, we isolated exosome-like vesicles from excretory/secretory products (ESP) of T. pisiformis cysticercus. The morphology and size of purified vesicles were confirmed by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). The components of proteins and miRNAs within these vesicles were identified by proteomic analysis and high-throughput small RNA sequencing. The biological function of targets of exosomal miRNAs was predicted by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Moreover, the expression of Th1- and Th2-type immune response associated cytokines in RAW264.7 macrophages were evaluated by qPCR and ELISA. We found that exosome-like vesicles were typical cup-shaped vesicles with diameters from 30 to 150 nm. A total of 87 proteins were identified by proteomic analysis, including proteins prominently associated with exosome-like vesicles biogenesis and vesicle trafficking. 41 known miRNAs and 18 novel miRNAs were identified in the exosome-like vesicles. Eleven selected miRNAs, including 7 known miRNAs (miR-71-5p, miR-10a-5p, miR-let-7-5p, miR-745-3p, miR-219-5p, miR-124-3p and miR-4989-3p) and 4 novel miRNAs (novel-mir-3, novel-mir-7, novel-mir-8 and novel-mir-11) were validated to exist in metacestiodes and exosome-like vesicles of T. pisiformis cysticercus by qPCR. The functions of most targets of exosomal miRNAs were mainly associated with signal transduction and the immune system. Additionally, T. pisiformis cysticercus-derived vesicles induced the production of IL-4, IL-6, IL-10, IL-13 and Arg-1, but downregulated the expression of IL-12, IFN-γ and iNOS in RAW264.7 macrophages. CONCLUSIONS: We demonstrated that proteins and miRNAs enclosed within exosome-like vesicles from T. pisiformis cysticercus have immunomodulatory functions. Furthermore, exosome-like vesicles were shown to induce the macrophage Th2-type immune response in vitro. Our study suggests that exosome-like vesicles play an important role in the interaction between cysticerci and their hosts.

Newly excysted juveniles of Fasciola gigantica trigger the release of water buffalo neutrophil extracellular traps in vitro.

Polymorphonuclear neutrophils (PMNs) are the most abundant leukocytes and are among the first line of immune system defense. PMNs can form neutrophil extracellular traps (NETs) in response to some pathogens. The release of NETs plays an important role in trapping and killing invading parasites. However, the effects of NETs on parasitic trematode infections remain unclear. In the present study, water buffalo NET formation, triggered by the newly excysted juveniles (NEJs) of Fasciola gigantica, was visualized by scanning electron microscopy. The major components of the structure of NETs were characterized by immunofluorescence. Viability of flukes incubated with water buffalo PMNs were examined under light microscopy. The results revealed that F. gigantic juveniles triggered PMN-mediated NETs. These NETs were confirmed to comprise the classic characteristics of NETs: DNA, histones, myeloperoxidase and neutrophil elastase. Although NETs were formed in response to viable larvae, the larvae were not killed in vitro. These results suggest that NET formation may serve as a mechanism to hamper the migration of large larvae to facilitate immune cells to kill them. This study demonstrates, for the first time, that parasitic trematode juveniles can trigger NET formation.

Relative safety of various spermatogenic stem cell purification methods for application in spermatogenic stem cell transplantation.

BACKGROUND: Spermatogonial stem cell (SSC) transplantation technology as a promising option for male fertility preservation has received increasing attention, along with efficient SSC purification technology as a necessary technical support; however, the safety of such application in patients with tumors remains controversial. METHODS: In this study, we used a green fluorescent protein mouse xenograft model of B cell acute lymphocytic leukemia. We isolated and purified SSCs from the testicular tissue of model mice using density gradient centrifugation, immune cell magnetic bead separation, and flow cytometry. The purified SSCs were transplanted into convoluted seminiferous tubules of the nude mice and C57BL/6 male mice subjected to busulfan. The development and proliferation of SSCs in the recipient testis were periodically tested, along with whether B cell acute lymphocytic leukemia was induced following SSC implantation. The genetic characteristics of the offspring obtained from natural mating were also observed. RESULTS: In testicular leukemia model mice, a large number of BALL cells infiltrated into the seminiferous tubule, spermatogenic cells, and sperm cells in the testis tissue decreased. After spermatogonial stem cell transplantation, the transplanted SSCs purified by immunomagnetic beads and flow cytometry methods colonized and proliferated extensively in the basement of the seminiferous tubules of mice; a large number of spermatogenic cells and sperm were found in recipient testicular tissue after 12 weeks of SSC transplantation. In leukemia detection in nude mice after transplantation in the three SSC purification groups, a large number of BALL cells could be detected in the blood of recipient mice 2-3 weeks after transplantation in the density gradient centrifugation group, but not in the blood of the flow cytometry sorting group and the immunomagnetic bead group after 16 weeks of observation. CONCLUSIONS: In this study, we confirmed that immunomagnetic beads and flow cytometry methods of purifying SSCs from the testicular tissue of the testicular leukemia mouse model could be safely applied to the SSC transplantation technology without concomitant tumor implantation. The results thus provide a theoretical basis for the application of tumor SSC cryopreservation for fertility preservation in patients with tumors.

[Rho/ROCK signaling pathway and anti-cryodamage ability of human sperm].

OBJECTIVE: To investigate the influence of the Rho/ROCK signaling pathway on the anti-cryodamage ability of human sperm and provide some theoretical evidence for the development of high-efficiency semen cryoprotectants. METHODS: We collected semen samples from 25 healthy males, each divided into a fresh, a normal cryopreservation control and an Rho-inhibition group. Before and after freezing, we detected sperm motility, viability, membrane integrity, morphology, DNA fragmentation index (DFI), acrosomal enzyme activity (AEA) and mitochondrial membrane potential (MMP) and determined the expressions of RhoA and ROCK proteins in the sperm by immunofluorescence staining. RESULTS: Compared with the normal cryopreservation control, the frozen-thawed sperm of the Rho-inhibition group showed significantly increased sperm motility （ ï¼»51.20 ± 7.70ï¼½% vs ï¼»57.50 ± 6.83ï¼½%, P = 0.002), survival rate ( ï¼»52.87 ± 5.07ï¼½% vs ï¼»60.24 ± 5.53ï¼½%, P = 0.001), membrane integrity (ï¼»59.78±5.56ï¼½% vs ï¼»67.10 ± 4.43ï¼½%, P = 0.001), percentage of morphologically normal sperm (ï¼»4.83 ± 1.11ï¼½% vs ï¼»7.46 ± 1.28ï¼½, P = 0.001) and MMP (56.30 ± 4.28 vs 63.11 ± 2.97, P = 0.001), but decreased DFI (ï¼»27.64 ± 6.64ï¼½% vs ï¼»18.87 ± 4.07ï¼½%, P = 0.001). There was no statistically significant difference in the AEA of the frozen-thawed sperm between the control and Rho-inhibition groups (97.65 ± 9.31 vs 98.30 ± 11.33, P > 0.05). Immunofluorescence staining revealed extensive expressions of RhoA and ROCK proteins in the head and neck of the sperm. CONCLUSIONS: The Rho/ROCK signaling pathway plays a role in the cryodamage to human sperm, and inhibiting the activity of Rho/ROCK can significantly improve the ability of sperm to resist cryodamage.

[Construction of a mouse model of spermatogonial stem cell transplant recipient by high-temperature heat stress].

OBJECTIVE: To investigate the feasibility of constructing a mouse model of spermatogonial stem cell (SSC) transplant recipient by high-temperature heat stress. METHODS: Four-week-old C57BL/6 male mice and B6(Cg)-Tyrc-2J/J coat color gene homozygous mutant male mice were heat-treated at 43 ℃ for an hour in the incubator. The best transplantation time was determined by HE staining, immunohistochemistry and TUNEL and the SSCs were transplanted into the seminiferous tubules of the mice followed by regular observation of the proliferation, differentiation and spermiogenesis of the SSCs in the testis of the recipient mice. Then the recipients were mated with age-matched normal female mice and the epigenetic features of their offspring were observed. RESULTS: After 3－5 days of high-temperature heat stress, the spermatogenic cells in the testicular seminiferous tubules of the recipient mice showed obviously decreased layers, disordered and loose arrangement, massive deletion, significant apoptosis, reduced mesenchymal cells and increased autophagy, which were basically recovered in about 12 days. At 8 weeks after transplantation, the isolated and purified SSCs were differentiated into spermatogenic cells and sperm with genetic function in the testicular seminiferous tubules of the recipient mice, and normal offspring were reproduced after natural mating. CONCLUSIONS: High-temperature heat stress can be used as an efficient method for rapid construction of the mouse model of spermatogonial stem cell transplantation recipient.

A novel extradural nerve transfer technique by coaptation of C4 to C5 and C7 to C6 for treating isolated upper trunk avulsion of the brachial plexus.

The study aimed to demonstrate the feasibility of an extradural nerve anastomosis technique for the restoration of a C5 and C6 avulsion of the brachial plexus. Nine fresh frozen human cadavers were used. The diameters, sizes, and locations of the extradural spinal nerve roots were observed. The lengths of the extradural spinal nerve roots and the distance between the neighboring nerve root outlets were measured and compared in the cervical segments. In the spinal canal, the ventral and dorsal roots were separated by the dura and arachnoid. The ventral and dorsal roots of C7 had sufficient lengths to anastomose those of C6. The ventral and dorsal of C4 had enough length to be transferred to those of C5, respectively. The feasibility of this extradural nerve anastomosis technique for restoring C5 and C6 avulsion of the brachial plexus in human cadavers was demonstrated in our anatomical study.