Toripalimab plus axitinib versus sunitinib as first-line treatment for advanced renal cell carcinoma: RENOTORCH, a randomized, open-label, phase III study.

BACKGROUND: Immune checkpoint inhibitors in combination with tyrosine kinase inhibitors are standard treatments for advanced clear cell renal cell carcinoma (RCC). This phase III RENOTORCH study compared the efficacy and safety of toripalimab plus axitinib versus sunitinib for the first-line treatment of patients with intermediate-/poor-risk advanced RCC. PATIENTS AND METHODS: Patients with intermediate-/poor-risk unresectable or metastatic RCC were randomized in a ratio of 1 : 1 to receive toripalimab (240 mg intravenously once every 3 weeks) plus axitinib (5 mg orally twice daily) or sunitinib [50 mg orally once daily for 4 weeks (6-week cycle) or 2 weeks (3-week cycle)]. The primary endpoint was progression-free survival (PFS) assessed by an independent review committee (IRC). The secondary endpoints were investigator-assessed PFS, overall response rate (ORR), overall survival (OS), and safety. RESULTS: A total of 421 patients were randomized to receive toripalimab plus axitinib (n = 210) or sunitinib (n = 211). With a median follow-up of 14.6 months, toripalimab plus axitinib significantly reduced the risk of disease progression or death by 35% compared with sunitinib as assessed by an IRC [hazard ratio (HR) 0.65, 95% confidence interval (CI) 0.49-0.86; P = 0.0028]. The median PFS was 18.0 months in the toripalimab-axitinib group, whereas it was 9.8 months in the sunitinib group. The IRC-assessed ORR was significantly higher in the toripalimab-axitinib group compared with the sunitinib group (56.7% versus 30.8%; P < 0.0001). An OS trend favoring toripalimab plus axitinib was also observed (HR 0.61, 95% CI 0.40-0.92). Treatment-related grade ≥3 adverse events occurred in 61.5% of patients in the toripalimab-axitinib group and 58.6% of patients in the sunitinib group. CONCLUSION: In patients with previously untreated intermediate-/poor-risk advanced RCC, toripalimab plus axitinib provided significantly longer PFS and higher ORR than sunitinib and had a manageable safety profile TRIAL REGISTRATION: ClinicalTrials.gov NCT04394975.

Potential functions and causal associations of VPS29 in hepatocellular carcinoma: a bioinformatic and Mendelian randomization study.

OBJECTIVE: Vacuolar protein sorting-associated protein 29 (VPS29) plays a certain role in cancer, but its biological significance in hepatocellular carcinoma (HCC) has not been studied. We utilized bioinformatics and Mendelian randomization (MR) analysis to explore the potential function of the VPS29 gene in HCC, as well as the causal relationship between VPS29 protein and HCC. MATERIALS AND METHODS: We downloaded the raw data from TCGA, GEO, and IEU OpenGWAS databases. We used R software for data processing and analysis to explore the relationship between the VPS29 gene and the expression, prognosis, clinical features, methylation, immune microenvironment, tumor mutation burden, and drug sensitivity in HCC patients. Additionally, a two-sample Mendelian randomization analysis was conducted to investigate the causal relationship between the VPS29 protein and HCC. RESULTS: VPS29 was found to be overexpressed in various types of cancer, including HCC, and its elevated expression often predicts poor prognosis in HCC patients. Univariate and multivariate Cox analysis demonstrated that VPS29 was an independent prognostic factor in HCC patients. The ROC curve indicated that VPS29 has a high diagnostic value in HCC. There were differential expressions of VPS29 in various clinical feature subgroups. The expression of VPS29 was negatively correlated with methylation levels, and multiple methylation sites were identified in the promoter region, including cg20877181, cg03867797, cg10025392, cg21605021, which were associated with poorer overall survival (OS) at low methylation levels. VPS29 was associated with immune cell infiltration disorders, including CD8+ T cells, Eosinophils, Neutrophils, Tcm, NK CD56bright cells, TFH, Th2 cells, Th17 cells, etc. Drug sensitivity analysis showed that VPS29 could be indicative of treatment response to 10 common antineoplastic drugs in different expression subgroups. Inverse variance weighted (IVW) analysis revealed a significant increase in HCC risk associated with VPS29 [odds ratio (OR): 1.440; 95% confidence interval (CI): 1.195-1.736], and sensitivity analysis showed no heterogeneity or pleiotropy. CONCLUSIONS: VPS29 is a risk factor for the occurrence and progression of HCC and may serve as a molecular biomarker for the diagnosis and prognosis of HCC.

[Safety and efficacy of laparoscopic surgery in locally advanced gastric cancer patients with neoadjuvant chemotherapy combined with immunotherapy].

Objective: To investigate the safety and efficacy of laparoscopic surgery in locally advanced gastric cancer patients with neoadjuvant SOX chemotherapy combined with PD-1 inhibitor immunotherapy. Methods: Between November 2020 and April 2021, patients with locally advanced gastric cancer who were admitted to the Union Hospital of Tongji Medical College of Huazhong University of Science and Technology were prospectively enrolled in this study. Inclusion criteria were: (1) patients who signed the informed consent form voluntarily before participating in the study; (2) age ranging from 18 to 75 years; (3) patients staged preoperatively as cT3-4N+M0 by the TNM staging system; (4) Eastern Collaborative Oncology Group score of 0-1; (5) estimated survival of more than 6 months, with the possibility of performing R0 resection for curative purposes; (6) sufficient organ and bone marrow function within 7 days before enrollment; and (7) complete gastric D2 radical surgery. Exclusion criteria were: (1) history of anti-PD-1 or PD-L1 antibody therapy and chemotherapy; (2) treatment with corticosteroids or other immunosuppre- ssants within 14 days before enrollment; (3) active period of autoimmune disease or interstitial pneumonia; (4) history of other malignant tumors; (5) surgery performed within 28 days before enrollment; and (6) allergy to the drug ingredients of the study. Follow-up was conducted by outpatient and telephone methods. During preoperative SOX chemotherapy combined with PD-1 inhibitor immunotherapy, follow-up was conducted every 3 weeks to understand the occurrence of adverse reactions of the patients; follow-up was conducted once after 1 month of surgical treatment to understand the adverse reactions and survival of patients. Observation indicators were: (1) condition of enrolled patients; (2) reassessment after preoperative therapy and operation received (3) postoperative conditions and pathological results. Evaluation criteria were: (1) tumor staged according to the 8th edition of the American Joint Committee on Cancer (AJCC) TNM staging system; (2) tumor regression grading (TRG) of pathological results were evaluated with reference to AJCC standards; (3) treatment-related adverse reactions were evaluated according to version 5.0 of the Common Terminology Criteria for Adverse Events; (4) tumor response was evaluated by CT before and after treatment with RECIST V1.1 criteria; and (5) Clavien-Dindo complication grading system was used for postoperative complications assessment. Results: A total of 30 eligible patients were included. There were 25 males and 5 females with a median age of 60.5 (35-74) years. The primary tumor was located in the gastroesophageal junction in 12 cases, in the upper stomach in 8, in the middle stomach in 7, and in the lower stomach in 3. The preoperative clinical stage of 30 cases was III. Twenty-one patients experienced adverse reactions during neoadjuvant chemotherapy combined with immunotherapy, including four cases of CTCAE grade 3-4 adverse reactions resulting in bone marrow suppression and thoracic aortic thrombosis. All cases of adverse reactions were alleviated or disappeared after active symptomatic treatment. Among the 30 patients who underwent surgery, the time from chemotherapy combined with immunotherapy to surgery was 28 (23-49) days. All 30 patients underwent laparoscopic radical gastrectomy, of which 20 patients underwent laparoscopic-assisted radical gastric cancer resection; 10 patients underwent total gastrectomy for gastric cancer, combined with splenectomy in 1 case and cholecystectomy in 1 case. The surgery time was (239.9±67.0) min, intraoperative blood loss was 84 (10-400) ml, and the length of the incision was 7 (3-12) cm. The degree of adenocarcinoma was poorly differentiated in 18 cases, moderately differentiated in 12 cases, nerve invasion in 11 cases, and vascular invasion in 6 cases. The number lymph nodes that underwent dissection was 30 (17-58). The first of gas passage, the first postoperative defecation time, the postoperative liquid diet time, and the postoperative hospitalization time of 30 patients was 3 (2-6) d, 3 (2-13) d, 5 (3-12) d, and 10 (7-27) d, respectively. Postoperative complications occurred in 23 of 30 patients, including 7 cases of complications of Clavien-Dindo grade IIIa or above. Six patients improved after treatment and were discharged from hospital, while 1 patient died 27 days after surgery due to granulocyte deficiency, anemia, bilateral lung infection, and respiratory distress syndrome. The remaining 29 patients had no surgery-related morbidity or mortality within 30 days of discharge. Postoperative pathological examination showed TRG grades 0, 1, 2, and 3 in 8, 9, 4, and 9 cases, respectively, and the number of postoperative pathological TNM stages 0, I, II, and III was 8, 7, 8, and 7 cases, respectively. The pCR rate was 25.0% (8/32). Conclusion: Laparoscopic surgery after neoadjuvant SOX chemotherapy combined with PD-1 inhibitor immunotherapy for locally advanced gastric cancer is safe and feasible, with satisfactory short-term efficacy. Early detection and timely treatment of related complications are important.

Incidence and risk factors of female urinary incontinence: a 4-year longitudinal study among 24 985 adult women in China.

OBJECTIVE: To estimate the incidence of urinary incontinence (UI), including its subtypes stress UI (SUI), urgency UI (UUI) and mixed UI (MUI), and to examine risk factors for de novo SUI and UUI in Chinese women. DESIGN: Nationwide longitudinal study. SETTING: Six geographic regions of China. PARTICIPANTS: Women aged ≥20 years old were included using a multistage, stratified, cluster sampling method. METHODS: This study was conducted between May 2014 and March 2016, with follow up in 2018. Data on demographics, medical history, lifestyle and physiological and anthropometric information were collected. MAIN OUTCOME MEASUREMENTS: Incidence, rate ratio (RR). RESULTS: Analyses included 24 985 women (mean age 41.9 years).The follow-up response rate was 55.5%, median follow-up time was 3.7 years. The standardised incidences of UI, SUI, UUI and MUI were 21.2, 13.1, 3.0 and 5.1 per 1000 person-years, respectively. Risk factors for de novo SUI included delivery pattern (vaginal spontaneous delivery RR 2.12, 95% CI 1.62-2.78 and instrumental delivery RR 3.30, 95% CI 1.99-5.45), high body mass index (BMI) (overweight RR 1.52, 95% CI 1.33-1.74 and obesity RR 1.67, 95% CI 1.32-2.11), cigarette smoking (RR 1.54, 95% CI 1.12-2.12), chronic cough (RR 1.44, 95% CI 1.17-1.76), diabetes (RR 1.33, 95% CI 1.10-1.60) and older age (50-59 years RR 1.49, 95% CI 1.16-1.90 and 60-69 years RR 1.61, 95% CI 1.22-2.13).The risk factors significantly associated with de novo UUI were age (RR increased from 1.21, 95% CI 0.74-1.99, at 30-39 years to 6.3, 95% CI 3.85-10.30, at >70 years) and diabetes (RR 1.48, 95% CI 1.05-2.09). CONCLUSIONS: The incidence of female UI is 21.2 per 1000 person-years in China. Delivery (vaginal spontaneous delivery, instrumental delivery), high BMI, cigarette smoking, chronic cough, diabetes and older age were risk factors. TWEETABLE ABSTRACT: The incidence of female urinary incontinence was 21.2 per 1000 person-years in China. Delivery, BMI, diabetes and old age are risk factors.

Effects of replacing soybean meal with pumpkin seed cake and dried distillers grains with solubles on milk performance and antioxidant functions in dairy cows.

Pumpkin seed cake (PSC), a byproduct of pumpkin seed oil processing, is used in ruminant feed as a beneficial protein source. Experiments were conducted to evaluate PSC as a substitute for soybean meal in the diets of lactating cows based on performance, rumen fermentation, antioxidant function and nitrogen partitioning. Six multiparous lactating cows were used in a replicated 3 × 3 Latin square experiment with 27-day periods. The cows were randomly divided into three treatment groups: group (1) was fed a diet containing no PSC (0PSC), and groups (2) and (3) were fed diets in which soybean meal was replaced with PSC and dried distillers grains with solubles (DDGS) at levels of 50% (50PSC) and 100% (100PSC), respectively. The diets were isonitrogenous and contained identical roughage but different proportions of PSC and DDGS. Replacement of soybean meal with PSC and DDGS did not influence rumen degradation, milk performance, rumen fermentation, DM intake or apparent total tract digestibility, and nitrogen partitioning between milk, feces and urine did not differ in the animals fed the three diets. However, compared with a diet containing no PSC, the total antioxidant capacity (P < 0.05) and antioxidant enzymes (total superoxide dismutase, glutathione peroxidase and catalase) activities (P < 0.05) were increased in the animals that received the 50PSC and 100PSC diets. In contrast, addition of PSC significantly reduced concentrations of aspartate transaminase (P < 0.05), alkaline phosphatase (P < 0.05) and malondialdehyde (P < 0.05) in the plasma. These results demonstrate that PSC can be completely substituted for soybean meal in the diet of dairy cows without any negative impact on milk performance, rumen fermentation or apparent digestibility and that this dietary change improves antioxidant functions and blood parameters in dairy cows, indicating that PSC has the potential for use as a feed source for dairy cows.

Circ-ABCB10 acts as an oncogene in glioma cells via regulation of the miR-620/FABP5 axis.

OBJECTIVE: This study aims to investigate the biological function of circular RNA ABCB10 (circ-ABCB10) in regulating the progression of glioma and to study the possible underlying mechanisms. PATIENTS AND METHODS: The expression levels of circ-ABCB10, miR-620 and FABP5 mRNA in glioma tissues, normal surrounding tissues and glioma cell lines were measured by Real-time PCR (RT-PCR). Circ-ABCB10 was silenced by siRNA in glioma cell lines (U87, T98G). The proliferation, migration and invasion of glioma cells were measured by MTT, wound healing and transwell assays, respectively. The relationship between circ-ABCB10, miR-620 and FABP5 was tested by Dual-Luciferase assay. The expression of proteins was measured by Western blot. The cell cycle distribution and apoptosis were measured by flow cytometry. RESULTS: The expression levels of circ-ABCB10 and FABP5 in glioma tissues and cells were significantly higher than those in their normal counterparts. Moreover, the expression of miR-620 was lower in glioma tissues. Silencing of circ-ABCB10 in glioma cells significantly inhibited the proliferation, migration and invasion of glioma cells. Moreover, downregulation of circ-ABCB10 induced cell cycle arrest and apoptosis in glioma cells. Furthermore, inhibition of miR-620 showed the opposite effects to silencing circ-ABCB10 on glioma cells. Dual-Luciferase reporter assays demonstrated that circ-ABCB10 could bind to miR-620 and that FABP5 was a direct target of miR-620. Western blot results showed that circ-ABCB10 could stabilize the expression of FABP5, while miR-620 decreased the expression of FABP5. Furthermore, overexpression of FABP5 abrogated the silencing effects of circ-ABCB10 in glioma cells. CONCLUSIONS: These data suggest that circ-ABCB10 affects glioma progression by regulating the miR-620/FABP5 axis, and circ-ABCB10 might be used as a potential target for the treatment of glioma.

[Correlation between fasting plasma glucose, HbA1c and DNA methylation in adult twins].

OBJECTIVE: To explore the cytidine-phosphate-guanosine (CPG) sites associated with fas-ting plasma glucose (FPG) and glycated haemoglobin (HbA1c) in twins. METHODS: In the study, 169 pairs of monozygotic twins were recruited in Qingdao, Zhejiang, Jiangsu, Sichuan and Heilongjiang in June to December of 2013 and June 2017 to October 2018. The methylation was detected by Illumina Infinium HumanMethylation450 BeadChip and Illumina Infinium MethylationEPIC BeadChip. According to the Linear Mixed Effect model (LME model), fasting plasma glucose and HbA1c were taken as the main effects, the methylation level (ß value) was taken as the dependent variable, continuous variables, such as age, body mass index (BMI), blood pressure, components of blood cells, surrogate variables generated by SVA, and categorical variables, such as gender, smoking and drinking status, hypoglycemic drugs taking, were included in the fixed effect model as covariates, and the identity numbers (ID) of the twins was included in the random effect model. The intercept was set as a random. Regression analysis was carried out to find out the CpG sites related to fasting blood glucose or HbA1c, respectively. RESULTS: In this study, 338 monozygotic twins (169 pairs) were included, with 412 459 CpG loci. Among them, 114 pairs were male, and 55 pairs were female, with an average age of (48.2±11.9) years. After adjustment of age, gender, BMI, blood pressure, smoking, drinking, blood cell composition, and other covariates, and multiple comparison test, 7 CpG sites (cg19693031, cg01538969, cg08501915, cg04816311, ch.8.1820050F, cg06721411, cg26608667) were found related to fasting blood glucose, 3 of which (cg08501915, ch.8.1820050f, cg26608667) were the newly found sites in this study; whereas 10 CpG sites (cg19693031, cg04816311, cg01538969, cg01339781, cg01676795, cg24667115, cg09029192, cg20697417, ch.4.1528651F, cg16097041) were found related to HbA1c, and 4 of which(cg01339781, cg24667115, cg20697417, and ch.4.1528651f) were new. We found that cg19693031 in TXNIP gene was the lowest P-value site in the association analysis between DNA methylation and fas-ting plasma glucose and HbA1c (PFPG=2.42×10-19, FDRFPG<0.001; PHbA1c=1.72×10-19, FDRHbA1c<0.001). CONCLUSION: In this twin study, we found new CpG sites related to fasting blood glucose and HbA1c, and provided some clues that partly revealed the potential mechanism of blood glucose metabolism in terms of DNA methylation, but it needed further verification in external larger samples.

High long non-coding LIFR-AS1 expression correlates with poor survival in gastric carcinoma.

OBJECTIVE: Abundant evidence has demonstrated that long non-coding RNAs (lncRNAs) play key roles in the development of human neoplasms. A novel cancer-related lncRNA, leukemia inhibitory factor receptor antisense RNA 1 (LIFR-AS1), has been reported to be under-expressed in breast cancer and associated with poor prognosis, but its significance in gastric cancer (GC) remains to be determined. Therefore, we assessed the prognostic and diagnostic value of LIFR-AS1 in GC. PATIENTS AND METHODS: Quantitative RT-PCR assay was used to detect the expression levels of LIFR-AS1 in GC tissues and adjacent normal tissues. The correlation between LIFR-AS1 expression and clinicopathological features was analyzed by Pearson's χ2-test. The disease-free survival and overall survival rates of GC patients were calculated by the Kaplan-Meier method. Cox regression analysis was used to assess factors related to survival. RESULTS: In this study, levels of LIFR-AS1 were significantly higher in GC tumor samples relative to adjacent normal tissue samples. A ROC analysis suggested LIFR-AS1 expression could be reliably used to differentiate between normal and GC tumor tissue. In addition, elevated LIFR-AS1 expression was positively correlated with more advanced and aggressive GC features, such as larger tumor size, lymphatic metastasis, and more advanced TNM stage. Survival analyses revealed that elevated LIFR-AS1 expression was correlated with worse overall survival and disease-free survival. Multivariate analysis further confirmed the relevance of LIFR-AS1 as an independent predictor of GC patient outcomes. CONCLUSIONS: In summary, these results indicate that the lncRNA LIFR-AS1 is a promising prognostic indicator in GC patients.

[Effect of Fei-Liu-Ping ointment combined with cyclophosphamide on lung cancer cell proliferation and acidic microenvironment].

OBJECTIVE: To observe the effects of Fei-Liu-Ping ointment and chemotherapy on mice with lung cancer, and to explore the inherent mechanism of action from the point of acidic microenvironment and apoptosis. METHODS: First of all, the Lewis lung cancer transplanted mouse model was established. Therefore, they were treated by Fei-Liu-Ping ointment, cyclophosphamide, Fei-Liu-Ping ointment + cyclophosphamide and the saline as control. All the groups' tumor size, tumor growth rate and food consumption were recorded. The mice were sacrificed and the tumors were took out after 15 days' interventions. Then lactate relative concentrations were detected with lactate kits and the protein expressions of glucose transporter 4 (GLUT4), hexokinase 1 (HK1), glucose-regulated protein 78 (GRP78), carbonic anhydrase-IX (CA-IX) were detected through immunohistochemical staining. Flow cytometry was adopted to detect the percentage of apoptotic tumor cells and regulatory T cells (Treg), and the expression of hypoxia-inducible factor-1α (HIF-1α), Bcl-2, Bax, Caspase-3, interleukin-2 (IL-2) were tested through western blot. RESULTS: The strongest inhibition effect and the lowest tumor growth rate was found in Fei-Liu-Ping ointment + cyclophosphamide group. There were significant differences between Fei-Liu-Ping ointment + cyclophosphamide group and saline group(P<0.05). And the highest food consumption was found in Fei-Liu-Ping ointment + cyclophosphamide group while there were no significant differences between Fei-Liu-Ping ointment + cyclophosphamide group and saline group (P>0.05). Further molecular biological detections found that the lowest lactate level and regulatory T cells ratio were found in Fei-Liu-Ping ointment + cyclophosphamide group and these expressions of GLUT4, HK1, GRP78, CA-IX were suppressed. There were significant differences between Fei-Liu-Ping ointment+cyclophosphamide group and saline group (P<0.05). In addition, the Fei-Liu-Ping ointment + cyclophosphamide group's cell apoptosis increased significantly compared with saline group and there were significant differences on expressions of HIF-1α, Bcl-2, Bax, Caspase-3, IL-2 for this group compared with saline group. CONCLUSION: Chemotherapy and Fei-Liu-Ping ointment had the synergistic effect on inhibiting tumor growth and improving the general conditions of tumor-bearing mice. The effect was partly owed to the improvement on tissue hypoxia, the inhibition of HIF-1α expression and the regulations on its downstream proteins, such as GLUT4, HK1, GRP78, and CA-IX. And then all these alterations led to the modulation tumor acidic microenvironment, the induced tumor cells apoptosis and suppression of T cells to regulatory T cells differentiation.

Effect of miR-26a-5p on gastric cancer cell proliferation, migration and invasion by targeting COL10A1.

OBJECTIVE: This study explored the effect of miR-26a-5p on cell proliferation, migration, and invasion in gastric cancer by targeting COL10A1. MATERIALS AND METHODS: First, differentially expressed genes were identified from microarray GSE103236 data of human gastric cancer. Then, qRT-PCR was carried out to detect the expression levels of COL10A1 and miR-26a-5p in gastric cancer cells and normal cases. The CCK-8 method was used to test cell proliferation. The colony formation assay was performed for the examination of the cell colony-forming ability, and transwell was applied for the detection of cell migration and invasion. Subsequently, the targeted relationship between miR-26a-5p and COL10A1 was identified by bioinformatics methods and further verified by Dual-Luciferase assay. The rescue experiment was finally conducted to validate the miR-26a-5p-dependent mechanism on cell proliferation, migration, and invasion via targeting COL10A1. RESULTS: COL10A1 was found to be highly expressed in gastric cancer cells, while miR-26a-5p was poorly expressed. Silencing COL10A1 inhibited cell proliferation, migration, and invasion in gastric cancer. Besides, miR-26a-5p could function on gastric cancer cells by reducing COL10A1. As well, the rescue experiment suggested that the down-regulation of COL10A1 could reverse the inhibitory effect of miR-26a-5p on gastric cancer cells. CONCLUSIONS: Collectively, miR-26a-5p can potentiate proliferation, migration, and invasion of gastric cancer cells by targeting COL10A1.

LncRNA NEAT1 promotes cardiac hypertrophy through microRNA-19a-3p/SMYD2 axis.

OBJECTIVE: The role of NEAT1 in cancers has been demonstrated. But the role of NEAT1 in cardiac hypertrophy still remains unknown. This study aimed to elucidate the specific function of long non-coding RNA (lncRNA) NEAT1 in cardiac hypertrophy and its underlying mechanism. PATIENTS AND METHODS: In this study, the in vivo and in vitro cardiac hypertrophy models were constructed by transverse aortic coarctation (TAC) procedure in rats and phenylephrine (PE) induction in primary cardiomyocytes, respectively. The expression levels of NEAT1, microRNA-19a-3p, SMYD2, and cardiac hypertrophic markers were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Cardiac hypertrophy was evaluated as calculating the surface area of hypertrophic cardiomyocyte by fluorescein isothiocyanate (FITC)-Phalloidin staining. Luciferase Reporter Gene Assay was conducted to detect the binding of NEAT1, SMYD2, and microRNA-19a-3p. RESULTS: The results showed that NEAT1 and SMYD2 were highly expressed in myocardium of rats with cardiac hypertrophy and PE-induced primary cardiomyocytes, whereas microRNA-19a-3p was lowly expressed. Besides, NEAT1 overexpression markedly upregulated levels of the cardiac hypertrophic markers. Moreover, FITC-Phalloidin staining also revealed hypertrophic cardiomyocytes overexpressing NEAT1. On the contrary, microRNA-19a-3p overexpression reduced the cardiomyocyte surface area and downregulated the levels of the cardiac hypertrophic markers. As luciferase reporter gene assay demonstrated, NEAT1 and SMYD2 could bind to microRNA-19a-3p. Finally, the gain-of-function experiments were designed to verify whether NEAT1 exerted its functions in cardiac hypertrophy by modulating SMYD2 and microRNA-19a-3p. Furthermore, both microRNA-19a-3p overexpression or SMYD2 knockdown could inhibit and reduce the cardiomyocyte surface area, and downregulate the levels of the cardiac hypertrophic markers. CONCLUSIONS: In summary, NEAT1 promotes the occurrence and progression of cardiac hypertrophy by upregulating SMYD2 by binding to microRNA-19a-3p.

Survival comparision of three-dimensional radiotherapy alone with concurrent chemoradiotherapy for non-surgical esophageal carcinoma.

PURPOSE: Radiotherapy is the main treatment method for patients with locally advanced, unresectable esophageal cancer. The aim of this study is to compare overall survival (OS) using 3D radiotherapy (3DRT) alone with concurrent chemoradiotherapy (CCRT) in 296 non-surgical esophageal carcinoma patients. PATENTS AND METHODS: Over 10 years, of the 480 patients with esophageal carcinoma treated with 3DRT with or without chemotherapy, 148 patients each comprised 3DRT and CCRT groups after propensity score matching. RESULTS: The 5- and 10-year OS (P=0.337) and PFS (P=0.715) rates for 3DRT alone were 22.0%, 14.4% and 26.1%, 23.2%, respectively, compared with 28.8%, 18.6% and 34.7%, 29.1% for CCRT, respectively. CCRT did not improve 5-year and 10-year OS or PFS in 60-70Gy group (OS: 27.5% and 25.2%; 17.9% and 17.0%, P=0.938; PFS: 38.3% and 31.8%; 31.9% and 27.8%, P=0.890) nor reduce 10-year hematogenous metastasis (31.7% and 28.3%, P=0.698). CCRT improved 5-year OS and PFS of 50.0-59.9Gy group (OS: 33.3% and 12.0%, P=0.029; PFS: 33.1% and 10.6%, P=0.081). For 3DRT, the 5-year OS and PFS rates were significantly better in the 60-70Gy group (P=0.017) compared with 50.0-59.9Gy group (P=0.002). For CCRT group, 5-year OS and PFS favored the 50.0-59.9Gy group, but the difference was insignificant. Major toxicities were greater with CCRT compared with 3DRT. CONCLUSION: For non-surgical esophageal carcinoma patients, 3DRT combined with CCRT was effective in prolonging both OS and PFS.

Klinefelter's syndrome with systemic lupus erythematosus and atrial fibrillation.

We describe a 65-year-old man who presented with arthralgia, reduced body hair and gynecomastia. He showed severe pancytopaenia. Laboratory examination revealed high follicle-stimulating hormone, low testosterone and oestradiol, elevated antinuclear antibodies, anti-dsDNA and ESR levels, as well as low complement levels. An electrocardiogram showed atrial fibrillation. Computed tomography and dual-energy x-ray absorptiometry showed pleural effusion and osteoporosis. Chromosome analysis revealed 47, XXY karyotype. The unifying diagnosis was therefore Klinefelter's syndrome (KS) with systemic lupus erythematosus (SLE), with manifestations of pancytopaenia, atrial fibrillation, serositis and osteoporosis. After immunosuppressive therapy, his physical condition and pancytopaenia improved. Sex hormones and gene escape from X chromosome inactivation may contribute to the pathogenesis of SLE. Clinicians should consider autoimmune processes when patients with KS present with pancytopaenia or additional features of a systemic autoimmune disorder.

Circ-CCDC66 accelerates proliferation and invasion of gastric cancer via binding to miRNA-1238-3p.

OBJECTIVE: The aim of this study was to examine the expression of circ-CCDC66 in gastric cancer (GC) tissues and cell lines, as well as its correlation with the prognosis of GC. Moreover, the regulatory effects of circ-CCDC66 on biological behaviors of GC cells and its molecular mechanism were explored. PATIENTS AND METHODS: The relative expression level of circ-CCDC66 in GC tissues and cell lines was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between the circ-CCDC66 level and overall survival of GC patients was analyzed as well. The potential influences of circ-CCDC66 on proliferative and invasive abilities of GC cells were evaluated through 5-Ethynyl-2'-deoxyuridine (EdU), colony formation and transwell assay, respectively. Meanwhile, the cell cycle progression and apoptosis of GC cells affected by circ-CCDC66 were determined. In addition, the direct target miRNA of circ-CCDC66 was predicted and verified by bioinformatics method and Dual-Luciferase reporter gene assay, respectively. RESULTS: Circ-CCDC66 was significantly up-regulated in GC tissues and cell lines. Up-regulation of circ-CCDC66 indicated markedly worse prognosis of GC patients. Transfection of circ-CCDC66-siRNA remarkably attenuated proliferative and invasive abilities of BGC-823 and MGC-803 cells. Besides, GC cells were arrested in the G0/G1 phase, and the apoptotic rate was remarkably elevated after circ-CCDC66 knockdown. The Dual-Luciferase reporter gene assay verified that circ-CCDC66 bind to miRNA-1238-3p by competing with LHX2 (LIM-homeobox domain 2). MiRNA-1238-3p was significantly down-regulated in GC cells, whereas LHX2 was up-regulated. Furthermore, overexpression of miRNA-1238-3p in GC cells markedly suppressed the LHX2 level. CONCLUSIONS: Circ-CCDC66 is highly expressed in GC tissues and cell lines. Knockdown of circ-CCDC66 attenuates proliferative and invasive abilities of GC cells. Our results indicate that circ-CCDC66/miRNA-1238-3p/LHX2 axis may be a promising target for GC treatment.

Clinical analysis of Chinese anti-low-density-lipoprotein-receptor-associated protein 4 antibodies in patients with myasthenia gravis.

BACKGROUND AND PURPOSE: Low-density-lipoprotein-receptor-associated protein 4 (LRP4) autoantibodies have recently been detected in myasthenia gravis (MG), but little is known about the clinical characteristics associated with this serological type. In this study, the clinical features of Chinese patients with anti-LRP4 antibody-positive MG were characterized. METHODS: A total of 2172 MG serum samples were collected from patients in various parts of China. An enzyme-linked immunosorbent assay was used to detect acetylcholine receptor (AChR) antibody and titin antibody, and cell-based assays were used to detect muscle-specific kinase antibody and LRP4 antibody. Clinical data for patients with MG were collected from different provinces in China. RESULTS: In total, 16 (0.8%) patients with LRP4-MG were found amongst 2172 total patients, including three patients with AChR/LRP4-MG. Additionally, 13 (2.9%) patients with LRP4-MG were found amongst 455 patients with double seronegative MG. The ratio of males to females for these 13 patients was 1:1.6, and 53.8% patients were children. A total of 91.7% of cases exhibited initial ocular involvement, and 58.3% of cases exhibited simple eye muscle involvement. Responses to acetylcholinesterase inhibitors and prednisone were observed. CONCLUSION: The expanded sample confirmed that the positive rate of LRP4 antibodies in China is lower than that in western countries. Our results highlighted the differences between LRP4-MG and other antibody groups. Children and female patients with LRP4-MG have a higher prevalence, often involving the ocular muscles and limb muscles. The clinical symptoms are mild, and satisfactory responses to treatment are often achieved.

[The application of three-dimensional printing technique combined with three-dimensional reconstruction in mandibular fracture].

Objective:To explore the value and significance of the combination of 3D printing and spiral CT three-dimensional reconstruction in the diagnosis and treatment of common mandibular fractures. Method:Forty patients with mandibular fracture were collected. They were randomly divided into traditional operation group and 3D printing combined with three-dimensional reconstruction operation group. The differences between the two groups were compared according to the operation time and the recovery of fracture. Result:The operation time and trauma of 3D printing combined with three-dimensional reconstruction group were significantly better than those of traditional operation group, and the former was better than the latter （P<0.05）. Conclusion: 3D printing combined with three-dimensional reconstruction operation method realized the concept of individualized, precise and minimally invasive comprehensive treatment, which is worthy of clinical promotion.

Identification and validation of novel microenvironment-based immune molecular subgroups of head and neck squamous cell carcinoma: implications for immunotherapy.

Background: Targeting the immune checkpoint pathway has demonstrated antitumor cytotoxicity in treatment-refractory head and neck squamous cell carcinoma (HNSC). To understand the molecular mechanisms underpinning its antitumor response, we characterized the immune landscape of HNSC by their tumor and stromal compartments to identify novel immune molecular subgroups. Patients and methods: A training cohort of 522 HNSC samples from the Cancer Genome Atlas profiled by RNA sequencing was analyzed. We separated gene expression patterns from tumor, stromal, and immune cell gene using a non-negative matrix factorization algorithm. We correlated the expression patterns with a set of immune-related gene signatures, potential immune biomarkers, and clinicopathological features. Six independent datasets containing 838 HNSC samples were used for validation. Results: Approximately 40% of HNSCs in the cohort (211/522) were identified to show enriched inflammatory response, enhanced cytolytic activity, and active interferon-γ signaling (all, P < 0.001). We named this new molecular class of tumors the Immune Class. Then we found it contained two distinct microenvironment-based subtypes, characterized by markers of active or exhausted immune response. The Exhausted Immune Class was characterized by enrichment of activated stroma and anti-inflammatory M2 macrophage signatures, WNT/transforming growth factor-ß signaling pathway activation and poor survival (all, P < 0.05). An enriched proinflammatory M1 macrophage signature, enhanced cytolytic activity, abundant tumor-infiltrating lymphocytes, high human papillomavirus (HPV) infection, and favorable prognosis were associated with Active Immune Class (all, P < 0.05). The robustness of these immune molecular subgroups was verified in the validation cohorts, and Active Immune Class showed potential response to programmed cell death-1 blockade (P = 0.01). Conclusions: This study revealed a novel Immune Class in HNSC; two subclasses characterized by active or exhausted immune responses were also identified. These findings provide new insights into tailoring immunotherapeutic strategies for different HNSC subgroups.

Clinical efficacy of sequential therapy with voriconazole on COPD patients in acute phase with pulmonary aspergillosis and effects on cytokines and pulmonary functions.

OBJECTIVE: To explore the clinical efficacy of sequential therapy with voriconazole on chronic obstructive pulmonary disease (COPD) patients in acute phase with pulmonary aspergillosis and its effects on cytokines and pulmonary functions. PATIENTS AND METHODS: A total of 110 COPD patients in acute phase with pulmonary aspergillosis who were admitted to the hospital between February 2015 and November 2016 were enrolled. We divided them randomly into two groups, i.e., the control group (n = 55) and the treatment group (n = 55). Patients in the control group took itraconazole capsules orally (200 mg/time, twice per day for three days followed by once per day). Patients in treatment group underwent sequential treatment with voriconazole through intravenous infusion at a dose of 5 mg/kg/time twice a day for 3 days followed by a dose of 4 mg/kg/time, twice a day for 8 days. Then, patients took voriconazole orally at a dose of 150 mL/time, twice a day for 6 days. Patients in two groups received the treatment for a total of 14 days. After treatment, we evaluated the levels of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and IL-8. The total lung capacity (TLC), diffusing capacity of the lung for carbon monoxide (DLco), and arterial oxygen saturation (SaO2), were measured as well. RESULTS: The total effectiveness rates of the treatment group and the control group were 83.63% and 61.82%. The differences had statistical significance (p < 0.01). After treatment, the incidence of chest pain, cough, sputum-coughing, hemoptysis, cyanosis, and dyspnea in the treatment group was significantly fewer than that in the control group (p < 0.05). TCL, DLco, and SaO2 in the two groups were significantly ameliorated by treatment (p < 0.05). The amelioration in the treatment group was more prominent than that in the control group (p < 0.05). The levels of TNF-α, IL-8, and IL-6 in the two groups were decreased dramatically by the treatments. The decrease in the treatment group was significantly lower than those in the control group (p < 0.05). Occurrence of adverse reactions in treatment group and control group were 8.33% and 6.25%, respectively; (p > 0.05). CONCLUSIONS: Sequential therapy with voriconazole exhibits promising clinical efficacy in COPD patients in acute phase with pulmonary aspergillosis. The treatment ameliorated the clinical symptoms and vital signs of patients significantly. It also improved the pulmonary functions and inhibited the inflammatory responses of patients with evident clinical efficacy.

Effects of gamma radiation on microbial, physicochemical, and structural properties of whey protein model system.

Gamma radiation has been used in food processing for many years, though it has certain effects on food components. Whey protein solutions (10%/30%, wt/vol) were treated with gamma radiation at various dosages (10-25 kGy) and evaluated for microbial changes in the solutions and physicochemical and structural changes of whey proteins. Whey protein solutions after gamma radiation showed substantially lower populations of all viable microorganisms than those of controls. The 10% whey protein solution treated at radiation of 20 or 25 kGy remained sterile for up to 4 wk at room temperature. Gamma radiation increased viscosity and turbidity and decreased soluble nitrogen of whey protein solutions compared to nonradiated control samples regardless of radiation dosage. Nonreducing sodium dodecyl sulfate-PAGE suggested that whey proteins under gamma radiation treatment formed aggregates with high molecular weights. Reducing sodium dodecyl sulfate-PAGE showed that disulfide bonds played a role in gamma radiation-induced whey protein cross-linking. Scanning and transmission electron microscopy micrographs exhibited large aggregates of whey proteins after gamma radiation treatment. Results suggested that gamma radiation could be applied to whey protein solution for purposes of reducing microbial counts and cross-linking protein molecules.

Comparison of CT radiogenomic and clinical characteristics between EGFR and KRAS mutations in lung adenocarcinomas.

AIM: To compare computed tomography (CT) radiogenomic and clinical characteristics between patients with epidermal growth factor receptor (EGFR) and Kirsten rat sarcoma viral oncogene (KRAS) mutations in lung adenocarcinomas. MATERIALS AND METHODS: This study was a retrospective analysis of patients with histopathologically confirmed lung adenocarcinoma, who had complete clinical and imaging data, and were tested for EGFR and KRAS mutations. Of the 313 included patients, 116 had effective EGFR mutations (EGFR group), 31 had KRAS mutations (KRAS group), and 166 had no EGFR or KRAS mutations (control group). Multivariate analysis was used to evaluate CT imaging features and clinical data. RESULTS: Multivariate analysis showed that significant variables between the EGFR and control groups were spiculation (odds ratio [OR] 2.70, 95% confidence interval [CI]: 1.54-4.75, p=0.001), and multiple small metastatic nodules (OR=7.52, 95% CI: 1.44-39.17, p=0.017). Significant variables between the KRAS and the control group were multiple small metastatic nodules (OR=7.65, 95% CI: 1.18-49.50, p=0.033). CONCLUSIONS: Patients with EGFR or KRAS mutations are prone to multiple metastases in both lungs. In addition, effective EGFR mutations mostly occurred in patients with multiple spiculations.