[New progress in the first-line treatment of advanced hepatocellular carcinoma].

Hepatocellular carcinoma is a kind of cancer with a strong invasion, a high incidence rate and mortality, and a poor prognosis. At the time of diagnosis, most patients are already in the advanced stages of a tumor and have lost the chance for radical surgical treatment. Advanced hepatocellular carcinoma treatment has a gradual transition from systemic chemotherapy to targeted therapy, immunotherapy, and combination therapy, especially immune checkpoint inhibitor-based immunotherapy combination therapy, such as combination with bevacizumab monoclonal antibodies and other drugs, or combination with TACE, HAIC, radiotherapy, ablation, and other treatment methods. Combination therapy has significant synergistic effects and thus has already become a future treatment trend for hepatocellular carcinoma. An immunotherapy-based combination therapy plan will run through the whole process of systemic therapy, which is expected to bring better survival benefits to patients with hepatocellular carcinoma. This article reviews the latest research progress in aspects of the first-line treatment of advanced hepatocellular carcinoma.

[Overexpression of tuftelin and KLF-5 and its clinicopathological features in hepatitis B virus-related hepatocellular carcinoma].

Objective: To analyze and evaluate the expressions and clinical value of tuftelin (TUFT1) and Krüppel-like factor 5 (KLF5) in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) tissues. Method: KLF5 mRNA and TUFT1 mRNA transcriptional status in cancer and non-cancer groups were compared according to the Cancer Genome Atlas (TCGA) database. The differences and prognostic value between the groups were analyzed. Postoperative liver cancer and its paired pericancerous tissues, with the approval of the ethics committee, were collected to build tissue chips. The expression of KLF5 and TUFT1 and their intracellular localization were verified by immunohistochemistry. Tissue expression and clinicopathological characteristics were analyzed by immunoblotting. SPSS software was used to analyze the relationship between SPSS and patient prognosis. Results: The transcription level of TUFT1 or KLF5 mRNA was significantly higher in the HCC group than the non-cancer group (P < 0.001), according to TCGA data. Immunohistochemistry and Western blotting examination confirmed the overexpression of TUFT1 and KLF5 in human HCC tissues, which were mainly localized in the cytoplasm and cell membrane. The positivity rates of TUFT1 and KLF5 were 87.1% ( χ(2) = 18.563, P < 0.001) and 95.2% ( χ(2) = 96.435, P < 0.001) in HCC tissues, and both were significantly higher than those in the adjacent group. The expression intensity was higher in stage III-IV than stage I-II of the International Union Against Cancer standard (P < 0.01). The clinicopathological features showed that the abnormalities of the two were significantly related to HBV infection, tumor size, extrahepatic metastasis, TNM stage, and ascites. Univariate analysis was related to tumor size, HBV infection, and survival. Multivariate analysis was an independent prognostic factor for patients with HCC. Conclusion: TUFT1 and KLF5 may both be novel markers possessing clinical value in the diagnosis and prognosis of HBV-related HCC.

Regulation of protein O-GlcNAcylation by circadian, metabolic, and cellular signals.

O-linked ß-N-acetylglucosamine (O-GlcNAcylation) is a dynamic post-translational modification that regulates thousands of proteins and almost all cellular processes. Aberrant O-GlcNAcylation has been associated with numerous diseases, including cancer, neurodegenerative diseases, cardiovascular diseases, and type 2 diabetes. O-GlcNAcylation is highly nutrient-sensitive since it is dependent on UDP-GlcNAc, the end product of the hexosamine biosynthetic pathway (HBP). We previously observed daily rhythmicity of protein O-GlcNAcylation in a Drosophila model that is sensitive to the timing of food consumption. We showed that the circadian clock is pivotal in regulating daily O-GlcNAcylation rhythms given its control of the feeding-fasting cycle and hence nutrient availability. Interestingly, we reported that the circadian clock also modulates daily O-GlcNAcylation rhythm by regulating molecular mechanisms beyond the regulation of food consumption time. A large body of work now indicates that O-GlcNAcylation is likely a generalized cellular status effector as it responds to various cellular signals and conditions, such as ER stress, apoptosis, and infection. In this review, we summarize the metabolic regulation of protein O-GlcNAcylation through nutrient availability, HBP enzymes, and O-GlcNAc processing enzymes. We discuss the emerging roles of circadian clocks in regulating daily O-GlcNAcylation rhythm. Finally, we provide an overview of other cellular signals or conditions that impact O-GlcNAcylation. Many of these cellular pathways are themselves regulated by the clock and/or metabolism. Our review highlights the importance of maintaining optimal O-GlcNAc rhythm by restricting eating activity to the active period under physiological conditions and provides insights into potential therapeutic targets of O-GlcNAc homeostasis under pathological conditions.

Nomogram Based on Clinical and Radiomics Data for Predicting Radiation-induced Temporal Lobe Injury in Patients with Non-metastatic Stage T4 Nasopharyngeal Carcinoma.

AIMS: To use pre-treatment magnetic resonance imaging-based radiomics data with clinical data to predict radiation-induced temporal lobe injury (RTLI) in nasopharyngeal carcinoma (NPC) patients with stage T4/N0-3/M0 within 5 years after radiotherapy. MATERIALS AND METHODS: This study retrospectively examined 98 patients (198 temporal lobes) with stage T4/N0-3/M0 NPC. Participants were enrolled into a training cohort or a validation cohort in a ratio of 7:3. Radiomics features were extracted from pre-treatment magnetic resonance imaging that were T1-and T2-weighted. Spearman rank correlation, the t-test and the least absolute shrinkage and selection operator (LASSO) algorithm were used to select significant radiomics features; machine-learning models were used to generate radiomics signatures (Rad-Scores). Rad-Scores and clinical factors were integrated into a nomogram for prediction of RTLI. Nomogram discrimination was evaluated using receiver operating characteristic analysis and clinical benefits were evaluated using decision curve analysis. RESULTS: Participants were enrolled into a training cohort (n = 139) or a validation cohort (n = 59). In total, 3568 radiomics features were initially extracted from T1-and T2-weighted images. Age, Dmax, D1cc and 16 stable radiomics features (six from T1-weighted and 10 from T2-weighted images) were identified as independent predictive factors. A greater Rad-Score was associated with a greater risk of RTLI. The nomogram showed good discrimination, with a C-index of 0.85 (95% confidence interval 0.79-0.92) in the training cohort and 0.82 (95% confidence interval 0.71-0.92) in the validation cohort. CONCLUSION: We developed models for the prediction of RTLI in patients with stage T4/N0-3/M0 NPC using pre-treatment radiomics data and clinical data. Nomograms from these pre-treatment data improved the prediction of RTLI. These results may allow the selection of patients for earlier clinical interventions.

[Activation of cGAS/STING signaling pathway and its immunological role in the progression of nonalcoholic fatty liver disease].

Today, nonalcoholic fatty liver disease remains the most dominant chronic liver disease. Cyclic guanosine monophosphate-adenosine monosphosphate synthase (cGAS) is a cytosolic DNA sensor that catalyzes the synthesis of cyclic guanosine monophosphate, activates stimulator of interferon genes (STING), and releases type-I interferon cytokines to trigger immune responses. Exogenous or endogenous DNA acts as a cGAS ligand to activate the cGAS-STING signaling pathway, which plays a role in hepatitis, nonalcoholic fatty liver disease, liver cancer and other diseases, and affects liver disease progression and metabolism through mechanisms such as autophagy. This article reviews the activation of cGAS-STING pathway and its molecular immunological role in nonalcoholic fatty liver disease progression.

[Effect of RBP2 gene silencing on proliferation, migration and invasion of ovarian epithelial cancer SKOV3/DDP cells and its mechanism].

Objective: To explore the effect of down-regulation of retinol binding protein 2 (RBP2) expression on the biological characteristics of ovarian cancer cells and its mechanism. Methods: Knockdown of RBP2 and cisplatin (DDP)-resistant ovarian cancer cell line SKOV3/DDP-RBP2i was established, the negative control group and blank control group were also set. Cell counting kit 8 (CCK-8) was used to detect the cell proliferation ability, flow cytometry was used to detect cell apoptosis, scratch test and Transwell invasion test were used to detect cell migration and invasion ability, real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) and western blot were used to detect the expressions of molecular markers related to epithelial-mesenchymal transition (EMT). The effect of RBP2 on the growth of ovarian cancer was verified through experiment of transplanted tumors in nude mice, and the relationships between RBP2 expression and tumor metastasis and patient prognosis were analyzed using the clinical data of ovarian cancer in TCGA database. Results: After down-regulating the expression of RBP2, the proliferation ability of SKOV3/DDP cell was significantly reduced. On the fifth day, the proliferation activities of SKOV3/DDP-RBP2i group, negative control group and blank control group were (56.67±4.16)%, (84.67±3.51) and (87.00±4.00)% respectively, with statistically significant difference (P<0.001). The apoptosis rate of SKOV3/DDP-RBP2i group was (14.19±1.50)%, higher than (8.77±0.75)% of the negative control group and (7.48±0.52)% of the blank control group (P<0.001). The number of invasive cells of SKOV3/DDP-RBP2i group was (55.20±2.39), lower than (82.60±5.18) and (80.80±7.26) of the negative control group and the blank control group, respectively (P<0.001). The scratch healing rate of SKOV3/DDP-RBP2i group was (28.47±2.72)%, lower than (50.58±4.06)% and (48.92±4.63)% of the negative control group and the blank control group, respectively (P<0.001). The mRNA and protein expressions of E-cadherin in the SKOV3/DDP-RBP2i group were higher than those in the negative control group (P=0.015, P<0.001) and the blank control group (P=0.006, P<0.001). The mRNA and protein expression of N-cadherin in SKOV3/DDP-RBP2i group were lower than those in the negative control group (P=0.012, P<0.001) and the blank control group (P=0.005, P<0.001). The mRNA and protein expressions of vimentin in SKOV3/DDP-RBP2i group were also lower than those in the negative control group (P=0.016, P=0.001) and the blank control group (P=0.011, P=0.001). Five weeks after the cells inoculated into the nude mice, the tumor volume of SKOV3/DDP-RBP2i group, negative control group and blank control group were statistically significant different. The tumor volume of SKOV3/DDP-RBP2i group was smaller than those of negative control group and blank control group (P=0.001). Bioinformatics analysis showed that the expression of RBP2 in patients with metastatic ovarian cancer was higher than that without metastasis (P=0.043), and the median overall survival of ovarian cancer patients with high RBP2 expression was 41 months, shorter than 69 months of low RBP2 expression patients (P<0.001). Conclusion: Downregulation of the expression of RBP2 in SKOV3/DDP cells can inhibit cell migration and invasion, and the mechanism may be related to the inhibition of EMT.

[Expression of METTL14 in epithelial ovarian cancer and the effect on cell proliferation, invasion and migration of A2780 and SKOV3 cells].

Objective: To study the expression of methyltransferase-like protein 14 (METTL14) in epithelial ovarian cancer and its clinical significance, and to explore the effect of METTL14 expression on the proliferation, invasion and migration of ovarian cancer cells. Methods: Immunohistochemistry (IHC) was used to detect METTL14 expression in tumor tissue samples, and analyze the relationships among METTL14 expression, clinicopathological factors, and prognosis in ovarian cancer. Lentiviral vectors and small interfering RNA (siRNA) were used to up-regulate and down-regulate the METTL14 expression in ovarian cancer cell lines A2780 and SKOV3, respectively. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was used to detect the N6-methyladenosine (m6A) content in ovarian cancer cells. Cell counting kit-8 (CCK-8), wound healing assay, and transwell assay were used to examine the function of METTL14 expression in the cells. Results: (1) The IHC score of METTL14 protein was 6.2±3.7 in 20 samples of ovarian cancer tissues and 3.3±2.5 in 15 samples of normal ovarian tissues, and the difference was statistically significant (t=-2.64, P=0.012). Among the patients who suffered from ovarian cancer, there were 69 cases with high expression of METTL14 protein (IHC score≥6), accounting for 57.0% (69/121), and the cases with low expression of METTL14 protein (IHC score<6) accounting for 43.0% (52/121). Compared with the patients with low expression of METTL14, the patients with high expression of METTL14 had later stages, higher rates of lymph node metastasis, abdominal metastasis, and more ascite amount. The differences were statistically significant (all P<0.05). The overall survival rate was significantly lower in patients with high METTL14 expression than the low expression (P=0.009). (2) LC-MS/MS data showed that the relative expression of m6A in A2780 and SKOV3 cells in the lentivirus (LV)-METTL14 group were 0.213±0.024 and 0.181±0.018, which were significantly higher than those in the LV-normal control (NC) group (0.109±0.022 and 0.128±0.020; all P<0.05). While the relative expression of m6A in A2780 and SKOV3 cells in the si-METTL14 group were 0.063±0.012 and 0.069±0.015, which were significantly lower than the expression in si-NC group of 0.108±0.014 and 0.121±0.014 (all P<0.05). CCK-8 assay showed that the absorbance values were significantly lower in the si-METTL14 group compared with the si-NC group at 36, 48, 60 hours (all P<0.05); while were significantly increased in the LV-METTL14 group compared with the LV-NC group at 48, 60 hours (all P<0.01). Scratch wound assays showed that the migration rate of the si-METTL14 group was lower than those of the si-NC group, while the LV-METTL14 group were higher than the LV-NC group by 24 hours, the differences were statistically significant (all P<0.01). Cell migration and invasion were detected by transwell migration and invasion assays. After cultivated for 24 hours, the invasion cell number and the migration cell number in the si-METTL14 group were less than those in the si-NC group. While the invasion cell number and the migration cell number in the LV-METTL14 group were more than those in the LV-NC group, respectively. The differences were statistically significant (all P<0.01). Conclusion: Patients with high METTL14 expression have a worse prognosis in ovarian cancer, which may increase the m6A modification of ovarian cancer cells and promote cells proliferation, invasion and migration.

[Abnormal expression of CD44 aggravates liver disease progression in patients with non-alcoholic fatty liver disease accompanied with hepatitis B virus replication].

Objective: To analyze the expression of CD44 in non-alcoholic fatty liver disease (NAFLD) accompanied with hepatitis B virus (HBV) infection and its clinical significance. Methods: Blood sample of hospitalized patients with NAFLD, chronic hepatitis B, cirrhosis, and healthy population (control) was collected. The study was approved by the hospital ethics committee. Serum CD44 level and clinopathological characteristics were analyzed quantitatively by enzyme-linked immunosorbent-assay. Flow cytometry was used to analyze the proportion of CD44(+)T lymphocytes in patients with NAFLD and chronic hepatitis B. NAFLD model was prepared with high-fat diet to verify the abnormal expression of CD44. Results: Compared with the healthy control group, the expression of serum CD44 in the cirrhosis group, chronic hepatitis B group and NAFLD group was increased, and the difference between the groups were statistically significant (P < 0.01). NAFLD patients graded as mild or severe group were equally accompanied by hepatocyte injury, abnormal blood glucose, lipid or CD44. In NAFLD patients accompanied with HBV infection, serum CD44 concentrations were significantly higher in HBsAg, HBeAg and HBV DNA positive group than HBsAg, HBeAg and HBV DNA negative group (P < 0.01). The proportion of CD44(+)T lymphocytes in peripheral blood of NAFLD and chronic hepatitis B group were 78.2% ± 16.3% and 68.5% ± 20.9%, respectively, and both groups (NAFLD and chronic hepatitis B) were significantly higher than the healthy control group (46.5% ± 20.5%) (P < 0.05). The high-fat diet model confirmed that in rat liver tissues the CD44 was overexpressed with fat deposition accompanied with liver cell damage, especially remarkable in liver tissues containing carcinogens. Conclusion: The abnormal expression of CD44 in patients with NAFLD may be related to the malignant transformation of HBV-related liver disease.

[Value of abnormal expression of Krüppel-like zinc-finger protein transcription factor 5 in the diagnosis and prognosis of liver cancer].

Objective: To explore the value of Krüppel-like factor 5 (KLF5), a family member of the zinc finger protein transcription factor, in the diagnosis and prognostic evaluation of hepatocellular carcinoma (HCC). Methods: Cancerous and non-cancerous tissues were collected from 126 cases after HCC surgery by self-matching method with microarray fabrication. Immunohistochemistry was used to analyze the expression of KLF5, clinicopathological characteristics and prognostic value. The sera of 222 cases with chronic liver disease were collected and their KLF5 levels were quantitatively determined by enzyme-linked immunosorbent assay (ELISA). Simultaneously, 40 normal human sera were used as controls to evaluate the value of abnormal KLF5 in the diagnosis and differentiation of benign and malignant liver diseases. T-test, Z-test and χ (2) test were performed on the data. Results: The positive expression rate of KLF5 in the HCC group was 95.2% (120/126), which was significantly higher than the non-cancerous group 38.9% (49/126; χ (2) = 14.385, P < 0.001). KLF5 expression was significantly correlated with TNM stage (stage I 35%, stage II 40%, stage III 74.4%, stage IV 78.1%), tumor size, alpha fetoprotein (AFP) concentration, portal vein embolism, HBV infection and 5-year survival rate. Univariate/multivariate analysis showed that KLF5 high expression was an independent predictor of HCC prognosis. The serum KLF5 level was significantly higher in HCC patients than liver cirrhosis, chronic hepatitis and normal control group (P < 0.001). With the serum KLF5 > 800 ng/ml and AFP > 25 µg/L as limit, the positive rates for HCC diagnosis were 90.48% and 73.81%, respectively, which were lower than the AFP specificity and false positive rate, and was helpful for the differential diagnosis of benign and malignant liver diseases. Conclusion: The overexpression of KLF5 in liver cancer tissues and blood is closely related to the HCC clinical stage and prognosis. Moreover, KLF5 analysis is helpful for HCC diagnosis and differential diagnosis.

[Investigation on high-frequency hearing loss of noise workers in an automobile factory].

Objective: To investigate the high-frequency hearing loss (HFHL) of workers in an automobile factory and its influencing factors, so as to provide a scientific basis for occupational health management of noise workers. Methods: From March to July 2019, 2647 workers from an automobile factory were selected as the research objects by cluster sampling method. The basic personal information (such as gender, age, education level, height, weight, etc.) , history of ear disease, the history of detonation, history of exposure to ototoxic drugs, history of exposure to occupational noise, etc., were investigated by questionnaire and occupational health examination. Chi-square test was used to compare the detection rates of HFHL in different populations, and multivariate logistic regression model was used to analyze the influencing factors of HFHL. Results: The noise level of 2647 individual workers ranged from 75.0 dB (A) to 92.0 dB (A) , with an average of (84.14±2.47) dB (A) . The detection rate of HFHL was 17.2% (456/2647) . There were significant differences in the the detection rate of HFHL among workers of different gender, age, educational level, working age, noise exposure level, diabetes history, detonation history, smoking and drinking workers (P<0.05) . Multivariate logistic regression analysis showed that male, age ≥30 years old, smoking and noise exposure were risk factors for HFHL (P<0.05) . Conclusion: The detection rate of HFHL in automobile factory workers is high. Gender, age, smoking and noise are the influencing factors of HFHL. It is necessary to strengthen the noise control in the workplace and guide workers to choose healthy lifestyle.

[Mechanism of T lymphocyte subsets in the malignant transformation of non-alcoholic fatty liver disease].

Nonalcoholic fatty liver disease is becoming the main cause of global liver disease-related morbidity and mortality. Notably, its pathological mechanism is complicated and not yet fully understood. Therefore, immune regulation is undoubtedly an important link in its pathogenesis, especially the change of T lymphocyte subsets. This article introduces the research progress of T lymphocytes involved in steatosis, inflammation, fibrosis, malignant transformation and immunotherapy.

[Expression of tuftelin protein and its clinical value in HBV-related liver cancer tissues].

Objective: To analyze the expression of tuftelin protein (TUFT1) and its clinical value in hepatocellular carcinoma (HCC)-related liver cancer tissues. Methods: The biological information data of TUFT1 mRNA expression in liver cancer and non-cancer tissues were analyzed from the TCGA and Oncomine database. After the approval of the ethics committee, the self-pairing method was used to collect the postoperative cancer and para-carcinoma tissues of 132 HCC cases hospitalized between January 2009 and December 2014. Tissue microarray and immunohistochemistry (IHC) were used to analyze the expression of TUFT1 in liver tissues. According to IHC staining, liver cancer was divided into high TUFT1 and low/no expression group. Combined with clinical data, the clinicopathological characteristics were statistically analyzed between and within the groups. The 5-year overall survival (OS) and disease-free survival (DFS) was analyzed by correlation analysis. Results: IHC staining showed that TUFT1 in cancer tissue was localized in the cytoplasm and cell membrane, and its positive expression rate was significantly higher in the liver cancer group (87.1%) than the para-carcinoma group (64.4%) (χ (2) = 18.563, P < 0.001). TUFT1 expression intensity in patients with liver cancer was significantly correlated with HBeAg positive (χ (2) = 4.080, P = 0.043), tumor size (χ (2) = 9.388, P = 0.002), vascular invasion (χ (2) = 14.885, P < 0.001), TNM stage (χ (2) = 13.516, P < 0.001) and ascites (χ (2) = 5.940, P = 0.015). TUFT1 high expression was negatively correlated with OS and DFS (P < 0.001). Conclusion: The overexpression of TUFT1 is closely related to HBV replication, vascular invasion and poor prognosis, and it is expected to become a useful marker for liver cancer diagnosis and prognosis.

[Phosphonate inhibits steatosis and lobular inflammation of non-alcoholic steatohepatitis through depleting macrophages].

Objective: To explore the role of macrophages in non-alcoholic steatohepatitis (NASH) in order to provide directions for the therapeutic target of metabolic liver disease. Methods: Twenty C57BL/6 wild-type male mice at 6-8 weeks were randomly divided into two groups: 5 in the control group, methionine-and choline-deficient diet (MCD); 15 in the experimental group, MCD diet + intraperitoneal injection of disodium chlorophosphonate liposomes (to clear macrophages). Mice were fed for 4 weeks to establish NASH model. Blood, liver and spleen were collected to analyze the body mass index, liver index, spleen index, and serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Non-alcoholic steatosis (NAS) activity score was evaluated by HE and Oil Red O staining. The relative expression level of F4/80 mRNA was compared by RT-PCR. Data comparison between groups was analyzed by t-test. Results: NASH model was successfully established by feeding the mice with MCD for four week. The expression of F4/80 mRNA (t = 4.167, P < 0.01), hepatic steatosis (t = 10.70, P < 0.05), interlobular inflammatory infiltration (t = 3.08, P < 0.05), and NAS score were decreased (t = 8.06, P < 0.05) in the experimental group. At the same time, ALT level [(817.00 ± 128.90) U/L vs. (231.20 ± 36.28) U/L, t = 5.71, P < 0.01], AST level [(1 211.00 ± 248.90) U/L vs. (505.30 ± 88.20) U/L, t = 3.32, P < 0.01] was decreased significantly. However, the spleen volume and spleen index of the experimental group were larger (0.24 ± 0.01 and 0.32 ± 0.02, t = 2.41, P < 0.05), and there was no significant effect on liver ballooning, body mass index and liver index. Conclusion: In NASH, phosphonate can consume macrophages to inhibit liver inflammation and protect the damaged liver.

Phosphatase and tensin homolog (PTEN) suppresses triacylglycerol accumulation and monounsaturated fatty acid synthesis in goat mammary epithelial cells.

Phosphatase and tensin homolog (PTEN) is a well-known tumor suppressor in nonruminants and regulates various cellular processes including growth through dephosphorylation of phosphoinositide substrates. Although studies with bovine mammary tissue suggested a role for PTEN during lactation, its potential role in lipid metabolism remains unknown. Objectives of the present study were to determine PTEN abundance in goat mammary tissue at 2 stages of lactation (n = 6 Xinong Saanen dairy goats per stage), and to use gene-silencing and adenoviral transfections in vitro with isolated goat mammary epithelial cells (GMEC) to evaluate the role of PTEN abundance of lipid metabolism-related genes. Abundance of PTEN decreased by 51.5% at peak lactation compared with the dry period. The PTEN was overexpressed in isolated GMEC through adenoviral transfection using an adenovirus system with Ad-GFP (recombinant adenovirus of green fluorescent protein) as control, and silenced via targeted small interfering RNA (siRNA) transfection with a scrambled small interfering RNA as a negative control. Cell culture was performed for 48 h before RNA extraction, triacylglycerol (TAG) analysis, and fatty acid analysis. Overexpression of PTEN downregulated abundance of acetyl-coenzyme A carboxylase α (ACACA), fatty acid synthase (FASN), sterol regulatory element binding transcription factor1 (SREBF1), stearoyl-coenzyme A desaturase 1 (SCD1), diacylglycerol acytransferase 1 (DGAT1), 1-acylglycerol-3-phosphate O-acyltransferase 6 (AGPAT6) coupled with an increase in patatin-like-phospholipase domain containing 2 (PNPLA2), hormone-sensitive lipase (LIPE), and carnitine palmitoyltransferase 1 ß (CPT1B). Furthermore, overexpressing PTEN in vitro resulted in a significant decrease in TAG concentration and concentration of C16:1. In contrast, interference of PTEN led to an opposite effect on lipid metabolism in GMEC. These changes suggested a shift from lipogenesis and esterification to lipolysis and fatty acid oxidation. Collectively, PTEN seems to play a role in monounsaturated fatty acids synthesis and lipid accumulation in GMEC.

[Hypoxia-inducible factor-1α mediates and regulates angiogenesis-related factors expression in hepatocellular carcinoma].

Objective: To investigate the expression of hypoxia-inducible factor (HIF)-1α, vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) during the progression of liver diseases and the molecular mechanism of HIF-1α in regulating the expression of angiogenic factors in hepatocellular carcinoma (HCC). Methods: Serum samples from hospitalized patients with liver cancer, liver cirrhosis, and chronic hepatitis were collected, and healthy people were used as controls. Mouse models of hepatocarcinogenesis were used to detect the dynamic expression of HIF-1α, VEGF and Ang-2. Enzyme-linked immunosorbent assay was used to quantitatively analyze the serum levels of HIF-1α, VEGF and Ang-2 in patients with liver disease and mice. HIF-1α-specific miRNA expression plasmids was constructed to transfect HepG2 human HCC cells. HIF-1α mRNA transcriptional interference effects were analyzed on biological behavior, VEGF and Ang-2 expression, and epithelial mesenchymal transformation (EMT) in human HCC cell line. The sample means of multiple groups were compared by analysis of variance and q test and the sample rate was compared by χ (2) test. Results: In patients with chronic liver disease, the serum expression of HIF-1α, VEGF and Ang-2 in the liver cancer group (145.6 ± 32.6) µg/L, (458.9 ± 125.3) µg/L and (42.9 ± 5.1) µg/L was significantly higher than the liver cirrhosis (P < 0.001) (79.5 ± 28.4) µg/L, (206.8 ± 56.8) µg/L and (26.2 ± 6.1) µg/L and chronic hepatitis group (60.1 ± 18.8) µg/L, (178.1 ± 85.4) µg/L and (21.8 ± 6.9) µg/L. In addition, HIF-1α was positively correlated with VEGF (r = 0.937, P < 0.001), HIF-1α and Ang-2 (r = 0.933, P < 0.001), and VEGF and Ang-2 (r = 0.910, P < 0.001). Mouse models of hepatocarcinogenesis confirmed that HIF-1α, VEGF and Ang-2 had progressively increased during the process of malignant transformation from normal hepatocytes, hepatocyte degeneration, and precancerous lesions to canceration. HIF-1α miRNA intervention plasmid had transformed HepG2 cells. Compared with the blank group, HIF-1α mRNA, HIF-1α, VEGF and Ang-2 were decreased by 88.1%, 59.8%, 54.0% and 36.0% at 72h, respectively. The expression level of EMT-related protein Snail (0.26 ± 0.02 and 0.67 ± 0.09, q = 6.75, P < 0.003), VIM (0.27±0.08 and 0.73±0.04, t = 10.35, P < 0.001) and Twist (0.24 ± 0.07 and 0.73 ± 0.02, q = 12.08, P < 0.001) was significantly reduced, but the expression level of E-cadherin (0.76 ± 0.08 and 0.27 ± 0.09, q = 7.05, P < 0.002) was significantly increased. Conclusion: HIF-1α mediates and regulates angiogenesis-related factors such as VEGF and Ang-2 expression in hepatocellular carcinoma. Furthermore, HIF-1α transcriptional interference can significantly affect the biological characteristics and EMT transformation of hepatocellular carcinoma cells.

[Surgical procedures for the correction and stabilization of pes planovalgus]. / Stabilisierung und Korrektur der medialen Säule im Rahmen der operativen Pes-planovalgus-Therapie.

The medial column of the foot is a relevant factor of the pathogenesis of pes planovalgus. Crucial anatomic structures are the tibiocalcaneonavicular ligament complex, the naviculocuneiform joints, including the ossa cuneiformia, and the first tarsometatarsal joint. A combination of bony and soft tissue reconstructive techniques must, therefore, be taken into account when treating pes planovalgus. The present article presents stabilizing and correcting surgical procedures for the medial column of the foot, including basic anatomy and biomechanics.

[Feasibility of near-infrared fluorescence imaging in assisting with the determination of the resection range of radiation intestinal injury].

Objective: To investigate the feasibility of near-infrared fluorescence imaging (NIRFI) to assist in determining the resection range of radiation intestinal injury (RII). Methods: A descriptive cohort study was conducted. Clinical data of 10 RII patients who presented intestinal obstruction and received operation with more than 100 cm of small intestine had been resected atGeneral Department of Jinling Hospital from October 2014 to January 2015 were retrospectively analyzed. The Novadaq SPY Intra-operative Imaging System was used in capturing and viewing fluorescent images. Firstly, the dense adhesion was mobilized and the obstructive intestine was fully freed under laparoscopy, then entering into abdomen from the corresponding incision. The surgeon determined the resection range according to the color of the intestinal serous layer of the diseased intestinal wall, the thickness of the intestinal wall, and the degree of swelling of the mesentery. Afterwards, intra-operative NIRFI was performed by intravenous injection of 2 ml indocyanine green (ICG) and the imaging results of the diseased intestinal arteriovenous phase were observed and recorded. The evaluation criteria for the final resection range were mainly based on the changes in mesenteric arterial phase imaging. In RII lesions, mesenteric vessels in mesenteric artery phase were disordered, and the comb-like distribution of normal mesenteric vessels completely disappeared. Only the clouded appearance in the intestinal wall was observed. Imaging results of the diseased intestinal tissue during the development phase and mesenteric vein phase were not significantly different from normal intestinal tissue. Intraoperative and postoperative conditions under NIRFI-assisted positioning, including the resection range, anastomosis site, operation-related complications, hospitalization time and cost were recorded. Data of abdominal contrast-enhanced CT and gastrointestinal angiography during 5 years of follow-up were collected to evaluate whether there was anastomotic stenosis or insufficient resection of diseased bowel. Results: Based on the imaging of mesenteric arterial phase of NIRFI, the median resection length of the small intestine was 185 (120-260) cm. After NIRFI imaging, only local lesion of ileum was excised in 6 patients, and jejunum-ileum anastomosis was performed to preserve ileocecal flap. No serious complications such as anastomotic leakage and anastomotic hemorrhage, or chronic intestinal failure such as short bowel syndrome occurred. The median hospitalization time was 32 (22-51) days, and the median hospitalization cost was 142 000 (90 000-175 000) RMB. The hospitalization time and cost were mainly used for the enteral and parenteral nutrition support treatment during the perioperative period. All the patients had normal oral diet and/or oral enteral nutrient. After 5 years of follow-up, no recurrence was found. Abdominal contrast-enhanced CT and gastrointestinal angiography showed no thickening of the intestinal wall or stenosis of the lumen. Conclusion: Mesenteric arterial phase imagingof NIRFI can help surgeons to determine the site and range of resection of RII lesions.

[Carcinoembryonic type specific markers and liver cancer immunotherapy].

Hepatocellular carcinoma (HCC) is a chronic inflammation derived from the background of hepatitis B and C virus (HBV and HCV) infection, chemical intoxicants, or non-alcoholic fatty liver disease. Cancerous liver cells can express and secrete a variety of relatively specific markers, such as carcinoembryonic type of alpha-fetoprotein (AFP), phosphatidylinositol-3 (Glypican-3, GPC-3), Wnt/ß-Catenin key molecule of signaling pathway Wnt3a and liver cancer specific GGT-II (HS-GGT), etc. Clinical analysis of carcinoembryonic markers not only contributes to diagnosis and prognosis of HCC, but may also be the target of HCC immunotherapy with a promising prospect of development and application. This article reviews the latest valuable advances in carcinoembryonic type specific molecular markers and liver cancer immunotherapy.

The knockdown of SNHG3 inhibits the progression of laryngeal squamous cell carcinoma by miR-340-5p/YAP1 axis and Wnt/ß-catenin pathway.

Laryngeal squamous cell carcinoma (LSCC) is a common malignancy of the head and neck. Long noncoding RNAs (lncRNAs) play essential roles in the development and treatment of LSCC. However, the role and regulatory mechanism of lncRNA small nucleolar RNA host gene 3 (SNHG3) in LSCC progression remain largely unknown. Twenty-five paired LSCC tissues and normal samples were collected. The expression levels of SNHG3, Yes-associated protein 1 (YAP1), and microRNA-340-5p (miR-340-5p) were measured via quantitative real-time polymerase chain reaction or western blot. Cell viability, apoptosis, and glycolysis were investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide, flow cytometry, and specific assay kit, respectively. The association between SNHG3 and miR-340-5p or miR-340-5p and YAP1 was assessed by dual-luciferase reporter assay. The expression of a protein involved in the Wnt/ß-catenin pathway was detected by western blot. The xenograft model was established to assess the anti-cancer role of SNHG3 inhibition in vivo. We found that the levels of SNHG3 and YAP1 were increased but the miR-340-5p expression was decreased in LSCC tissues and cells. The knockdown of SNHG3 or YAP1 inhibited cell viability and glycolysis but induced apoptosis in LSCC cells. Overexpression of YAP1 reversed the effect of SNHG3 knockdown on LSCC progression. SNHG3 could regulate YAP1 expression by competitively binding with miR-340-5p. Overexpression of miR-340-5p suppressed cell viability and glycolysis but promoted apoptosis in LSCC cells. Knockdown of SNHG3 repressed Wnt/ß-catenin pathway by regulating miR-340-5p and YAP1. The silencing of SNHG3 reduced LSCC xenograft tumor growth. In conclusion, knockdown of SNHG3 inhibited LSCC progression via inactivating Wnt/ß-catenin pathway by regulating the miR-340-5p/YAP1 axis.

[Clinical strategies for preservation of the exposed implant in chronic wounds and wound repair].

Objective: To explore the clinical strategies for preservation of the exposed implant in chronic wounds and wound repair. Methods: From January 2016 to January 2019, totally 8 patients (4 males and 4 females, aged 10 to 73 years) sustaining postoperative chronic wounds with exposed implants were admitted to the Fourth Medical Center of PLA General Hospital. There were 2 cases of abdominal patch exposure after abdominal trauma surgery, 2 cases of titanium plate exposure post craniocerebral surgery, 3 cases of internal fixator exposure post orthopedic surgery, and 1 case of cerebrospinal fluid drainage tube exposure after craniocerebral surgery. The wound exudate was collected for bacterial culture on admission. On the basis of glycemic control and correction of anemia and hypoproteinemia, thorough wound debridement was performed as soon as possible and the wound area after debridement ranged from 2.0 cm×0.5 cm to 6.0 cm×5.0 cm. The wounds of 4 patients were immediately closed after debridement, including 1 case by primary closure, 1 case by primary closure after local filling of platelet rich plasma gel, and 2 cases by local flap transplantation, with flap size of 10.0 cm×8.0 cm and 12.0 cm×8.0 cm, respectively. The donor sites of flaps were sutured directly and all the incisions were treated with continuous vacuum sealing drainage (VSD) after surgery. The other 4 patients were treated with continuous VSD after debridement to improve the wound bed. The wound of 1 case healed gradually, 1 case received direct wound suturing, and the wounds of 2 cases were repaired with thin split-thickness skin grafts from the thigh or the head. The results of bacterial culture of wound exudate on admission, wound healing post surgery, and follow-up were observed and recorded. Results: The bacterial culture of wound exudate on admission was positive in 6 patients, and 10 strains of bacteria were isolated with Staphylococcus epidermidis as the main pathogen. All the skin grafts or flaps of patients survived post surgery, with the incisions and wounds healed and all the implants preserved. After 1 to 3 years of follow-up, no recurrence of wound was found in any patient. Conclusions: The postoperative chronic wounds with exposed implants can be closed in primary stage by direct suturing or flap transplantation if it is clean enough on the basis of thorough debridement. The wounds with large defects or serious infection can be treated with continuous VSD firstly and then closed with direct suturing or skin grafting for delayed wound closure, thereby to reach the treatment goal of preserving the implants and repairing the wounds simultaneously.