[Doublecortin-like kinase 1 activates Hippo pathway to promote migration, invasion and proliferation of pancreatic cancer cells].

Objective: To explore the mechanism of Doublecortin-like kinase 1 (DCLK1) in promoting cell migration, invasion and proliferation in pancreatic cancer. Methods: The correlation between DCLK1 and Hippo pathway was analyzed using TCGA and GTEx databases and confirmed by fluorescence staining of pancreatic cancer tissue microarrays. At the cellular level, immunofluorescence staining of cell crawls and western blot assays were performed to clarify whether DCLK1 regulates yes associated protein1 (YAP1), a downstream effector of the Hippo pathway. Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) was used to analyze the expressions of YAP1 binding transcription factor TEA-DNA binding proteins (TEAD) and downstream malignant behavior-promoting molecules CYR61, EDN1, AREG, and CTGF. Transwell test of the DCLK1-overexpressing cells treated with the Hippo pathway inhibitor Verteporfin was used to examine whether the malignant behavior-promoting ability was blocked. Analysis of changes in the proliferation index of experimental cells used real-time label-free cells. Results: TCGA combined with GTEx data analysis showed that the expressions of DCLK1 and YAP1 molecules in pancreatic cancer tissues were significantly higher than those in adjacent tissues (P<0.05). Moreover, DCLK1was positively correlated with the expressions of many effectors in the Hippo pathway, including LATS1 (r=0.53, P<0.001), LATS2 (r=0.34, P<0.001), MOB1B (r=0.40, P<0.001). In addition, the tissue microarray of pancreatic cancer patients was stained with multicolor fluorescence, indicated that the high expression of DCLK1 in pancreatic cancer patients was accompanied by the up-regulated expression of YAP1. The expression of DCLK1 in pancreatic cancer cell lines was analyzed by the CCLE database. The results showed that the expression of DCLK1 in AsPC-1 and PANC-1 cells was low. Thus, we overexpressed DCLK1 in AsPC-1 and PANC-1 cell lines and found that DCLK1 overexpression in pancreatic cancer cell lines promoted YAP1 expression and accessible to the nucleus. In addition, DCLK1 up-regulated the expression of YAP1 binding transcription factor TEAD and increased the mRNA expression levels of downstream malignant behavior-promoting molecules. Finally, Verteporfin, an inhibitor of the Hippo pathway, could antagonize the cell's malignant behavior-promoting ability mediated by high expression of DCLK1. We found that the number of migrated cells with DCLK1 overexpressing AsPC-1 group was 68.33±7.09, which was significantly higher than 22.00±4.58 of DCLK1 overexpressing cells treated with Verteporfin (P<0.05). Similarly, the migration number of PANC-1 cells overexpressing DCLK1 was 65.66±8.73, which was significantly higher than 37.00±6.00 of the control group and 32.33±9.61 of Hippo pathway inhibitor-treated group (P<0.05). Meanwhile, the number of invasive cells in the DCLK1-overexpressed group was significantly higher than that in the DCLK1 wild-type group cells, while the Verteporfin-treated DCLK1-overexpressed cells showed a significant decrease. In addition, we monitored the cell proliferation index using the real-time cellular analysis (RTCA) assay, and the proliferation index of DCLK1-overexpressed AsPC-1 cells was 0.66±0.04, which was significantly higher than 0.38±0.01 of DCLK1 wild-type AsPC-1 cells (P<0.05) as well as 0.05±0.03 of DCLK1-overexpressed AsPC1 cells treated with Verteporfin (P<0.05). PANC-1 cells showed the same pattern, with a proliferation index of 0.77±0.04 for DCLK1-overexpressed PANC-1 cells, significantly higher than DCLK1-overexpressed PANC1 cells after Verteporfin treatment (0.14±0.05, P<0.05). Conclusion: The expression of DCLK1 is remarkably associated with the Hippo pathway, it promotes the migration, invasion, and proliferation of pancreatic cancer cells by activating the Hippo pathway.

[Association between serum lysophosphatidylcholine level and elderly health index in older people from longevity areas of Guangxi Province].

Objective: To investigate the relationship between serum lysophosphatidylcholine (LPC) level and the health index of the elderly. Methods: A total of 251 subjects were selected from the 2016 baseline survey of the Yongfu Longevity Cohort in Guangxi Province among whom 66, 63 and 122 were in the young and middle-aged group (≤59 years old), the young group (60-89 years old) and the longevity group (≥90 years old), respectively. Demographic data were collected and related indicators of height, weight, blood pressure and lipid metabolism were measured. The cognitive and physical functions of the elderly were assessed by the results of the simple mental state scale and the daily living activity scale to construct the health index of the elderly. The serum levels of LPC16∶0, LPC18∶0, LPC18∶1 and LPC18∶2 were determined by liquid chromatography-tandem mass spectrometry, and the differences among different ages and health status groups were compared. The logistic regression model was used to analyze the relationship between the serum LPC level and the health index of the elderly. Results: With the increase in age, the proportion of female subjects increased, and the rate of smoking and drinking decreased. BMI, TC, TG, LDL-C, diastolic blood pressure, and the four LPCs levels decreased with the increase of age, and systolic blood pressure levels increased with the increase of age (all P values<0.05). There was no significant difference in HDL-C levels among age groups (P>0.05). With the decline of health status in the elderly, serum levels of LPC16∶0, LPC18∶0, LPC18∶1 and LPC18∶2 showed a downward trend (all P values<0.001). After adjusting for age and gender, only LPC18∶0 was associated with the health status in old age [OR (95%CI): 0.48 (0.25-0.92)]. For every 1 standard deviation (16.87 nmol/L) increase in serum LPC18∶0 concentration, the risk of poor health status in old age decreased by 52%. Conclusion: Serum LPC18∶0 was associated with the health status in old age independent of age and sex.

[Research progress on main disease-related factors of healthy life expectancy].

International research on healthy life expectancy (HALE) focuses on inequality of socioeconomic status and individual natural attributes. With the acceleration of population ageing and the increase in average life expectancy, the extension of unhealthy life expectancy and the increase of social and economic burden caused by diseases have gradually attracted the attention of countries around the world. Therefore, the evaluation of disease factors affecting HALE is a meaningful direction in the future. This study introduces the development process and commonly used measurement methods of HALE. According to the definition of health from the Global Burden of Disease Study and World Health Organization, physical and mental diseases such as cardiovascular and cerebrovascular diseases, chronic respiratory diseases, diabetes, malignant tumors and depression were selected to summarize the impact of these diseases and pre-disease states on HALE. It is expected to provide a theoretical basis for the formulation of relevant public health policies and the improvement of quality of life in China.

Comparison of amino acid digestibility and its additivity determined with slaughter or cecectomy method for yellow-feather chicken.

The objective of this experiment was to compare the slaughter and cecectomy methods to determine amino acid (AA) digestibility of corn and soybean meal and their additivity in a corn-soybean meal diet. A completely randomized design was adopted to determine endogenous AA losses (EAAL) and AA digestibility in each of corn, soybean meal, and a corn-soybean meal diet using either slaughter or cecectomy methods. Each treatment contained 6 replicates with 3 chickens per replicate. The endogenous loss (EL) of histidine and glycine was lower and the EL of methionine and phenylalanine was greater when determined by slaughter vs. cecectomy (P < 0.05). The EL of arginine, isoleucine, leucine, lysine, methionine, phenylalanine, valine, alanine, aspartic acid, glutamic acid, and serine determined by slaughter were 1.2 to 3.2 times of those from cecectomy. The standard error (SE) of EL of 14 AA (excluding histidine and glycine) obtained by slaughter method was 2.1 to 9.6 times of those by cecectomy method. The apparent and standardized digestibility was not affected by methods for most AA except apparent digestibility of methionine, phenylalanine and glycine, and standardized digestibility of glycine in corn. The apparent and standardized digestibility of most AA except apparent digestibility of glycine and standardized digestibility of lysine, cysteine and glycine were less for slaughter versus cecectomy methods in soybean meal (P < 0.05). Using slaughter method resulted in reduced apparent digestibility of 15 AA (except glycine) and reduced standardized digestibility of 7 AA (arginine, isoleucine, leucine, valine, aspartic acid, glutamic acid, and proline) relative to cecectomy method (P < 0.05), but the standardized digestibility of glycine was greater when determined by slaughter vs. cecectomy methods in corn-soybean meal diet (P < 0.05). The mean value of SE of 16 AA digestibility in slaughter method was 2.9 times of that by cecectomy method. The apparent digestibility of 2 and 9 of 16 AA and the standardized digestibility of 15 and 7 of 16 AA were additive when using slaughter and cecectomy determinations, respectively. In conclusion, compared to the slaughter method, cecectomy method had less SE and EAAL but greater apparent digestibility of methionine and phenylalanine in corn, and the apparent digestibility of 15 AA (except glycine) in soybean meal and corn-soybean meal diet. Additivity in apparent and standardized AA digestibility was more inconsistent when determined with slaughter vs. cecectomy methods. These findings suggest that the cecectomy method is more suitable than the slaughter method to determine the digestibility of AA.

[Analysis of clinical features and surgical outcomes of petrous bone cholesteatomas].

Objective: To analyze the clinical features and surgical outcomes of petrous bone cholesteatomas (PBCs). Methods: Data from 39 PBCs patients treated in the Department of Otorhinolaryngology, Xijing Hospital from September 2011 to December 2017 were reviewed retrospectively, including 23 males, 16 femals, aged 12-71 years old, with the median age of 37. Clinical classifications, surgical methods, facial and hearing function, and intraoperative and postoperative complications were made summary analysis. Results: In this study, five patients were congenital PBCs and 34 patients were acquired PBCs. The common clinical symptoms were hearing loss (100%, 39/39), ear discharge/pus (89.7%, 35/39) and facial paralysis (46.2%, 18/39). According to Sanna's classification, 14 cases were supralabyrinthine, including three cases underwent transcochlear (TC) approach, six cases underwent transotic (TO) approach and five underwent translabyrinthine (TL) approach. 10 cases were infralabyrinthine, including eight cases underwent subtotal petrosectomy, one case underwent TO approach and one underwent TL approach.10 cases were massive, including seven cases underwent TC approach, three cases underwent TO approach. Five cases were infralabyrinthine-apical, including two cases underwent TC approach, two cases underwent TO approach, and one case underwent endoscope assisted infratemporal fossa type B. The degree of facial nerve (FN) dysfunction from high to low was massive (6/10), supralabyrinthine (8/14), infralabyrinthine-apical (2/5) and infralabyrinthine (2/10). 19 cases involved in facial nerve operation, three cases underwent FN decompression, four cases underwent FN rerouting, four cases underwent nerve grafting, and one case underwent facial-hypoglossal anastomosis. Preoperative FN involvement in 18 cases, and the FN function was improved in 14 cases after surgery. The improved rate of postoperative FN function was 77.8%. The bone conducted hearing retained 50.0% (14/28) postoperatively. Five cases with cerebrospinal fluid leak were managed by inserting free muscle plugs and cavity obliteration. Two cases with the cholesteatomas matrix involved the sigmoid sinus and the jugular bulb, and occlusion of the sigmoid sinus was performed. Postoperatively, two patients presented with synkinesis. The patients were followed up for 40 to 115 months, and there was no recurrence. Conclusions: There are no specific clinical manifestations for PBCs, thus, it is difficult in early diagnosis and treatment. According to Sanna's classification, preoperative FN and hearing function, the best surgical approach should be selected with minimal recurrences and perioperative morbidity.

[Assessment of cognitive function of the elderly by serum metabolites of brain-gut axis].

Objective: To investigate the feasibility of assessing cognitive function of the elderly by serum metabolites of brain-gut axis. Methods: Convenience sampling was used to select 100 and 60 participants from the healthy population cohort and microecological balance cohort of the longevity population in Guangxi, to constitute subset of healthy population and longevity population, respectively. A questionnaire was used to investigate the demographic characteristics of the subjects, 2-5 ml of fasting venous blood was collected from the subjects, and the serum untargeted metabolomics was determined by liquid chromatography tandem mass spectrometry. The biomarkers related to the brain-gut axis were collected through literature retrieval, and the results were intersected with the untargeted metabolites and annotated. Spearman correlation analysis was used to screen serum metabolites of brain-gut axis associated with aging, and multiple linear regression method was used to construct biological age model. The mini mental status examination was used to evaluate the cognitive function of longevity population subsets. The differences of biological age and chronological age of longevity population subsets with different cognitive function were compared. Results: The M (Q1, Q3) of subset of healthy population and longevity population were 64 (38, 72) and 97 (95, 99) years old, respectively, and there were 50 (50.0%) and 44 (73.3%) females, respectively. Nine serum metabolites of brain-gut axis were obtained by initial screening, which were propionic acid, glutamic acid, γ-aminobutyric acid (GABA), lactic acid, 5-hydroxytryptamine (5-HT), tryptophan, trimethylamine oxide, dopamine and canine urea. Spearman correlation analysis showed that glutamic acid and dopamine were positively correlated with aging (r values were 0.208 and 0.524, respectively, all P values<0.05), and tryptophan, 5-HT and GABA were negatively correlated with aging (r values were -0.308, -0.533 and -0.213, respectively, all P values<0.05). The biological age model was constructed as: y=49.81-1.18×10-5× GABA-1.82×10-4×5-HT+1.99×10-3×dopamine+1.65×10-6×glutamic acid -2.04×10-6×tryptophan+2.36×gender, where y was the biological age (years), the items on the right were the intercept item, the relative concentration of each metabolite, and gender (male=1, female=2). The coefficient of determination of model was 0.50 (P<0.001). The M (Q1, Q3) of the chronological age of the subset of longevity population with poor, moderate and good cognitive function were 97 (94, 100), 97 (93, 101) and 96 (94, 101) years old, respectively, and there was no statistical significance in pairwise comparison (all P values>0.05). The M (Q1, Q3) of the biological age of the subjects with better cognitive function was 51 (38, 54) years old, which was lower than that of the subjects with poor cognitive function [57 (47, 61)] (P=0.040). Conclusion: The biological age model can be constructed based on serum metabolites of brain-gut axis and used to evaluate the cognitive function of the elderly.

Spatiotemporal changes of CT manifestations in 110 patients with COVID-19 pneumonia.

OBJECTIVE: The aim of the study was to analyze spatiotemporal changes of CT manifestations in patients with COVID-19 pneumonia. PATIENTS AND METHODS: In this retrospective review, 110 patients with confirmed COVID-19 by RT-PCR form February 16, 2020, to March 28, 2020 were included. A total of 449 CT scans were reviewed. We analyze the type and distribution of lung abnormalities, and CT general assessment and lesion area statistics were performed. Patients were divided into mild, moderate, and severe disease based on Chinese guidelines: mild (patients with minimal symptoms, CT scans showed no pneumonia or a small area of pneumonia infection), moderate (different extent of clinical manifestations and CT scans showed multiple pneumonia infections in both lungs), severe disease (respiratory distress, CT scans lesion area exceeds 50%, and the lesion contains consolidation). The proportion of patients with mild, moderate and severe diseases was counted. RESULTS: The CT score and the area involved reached a peak (median 10) on illness days 7-12, and then, continued to be at a high level. The main abnormal pattern after symptoms appeared GGO (36/94 [36%] to 40/65 [62%] in different periods). The proportion of mixed reached its peak on illness days 13-18 (36/93 [39%]). Pure GGO was the most common subtype of GGO (24 of 60 CT scans [40%] to 23 of 33 CT scans [70%]) after symptoms onset. The ratio of GGO with irregular lines and interfaces peaked on illness days 7-12 (6/34 [18%]). The lesions are mainly distributed on both sides and under the pleura. 76/84 (90%) of discharged patients had residual lesions on the final CT scans. 4 confirmed patients' CT scans did not show lesions (on illness days 1-24 days). There were 47 mild cases (42.7%), 46 moderate cases (41.8%), and 7 severe cases (6.3%). CONCLUSIONS: The degree of lung abnormality on the CT of the patients reached the peak on the 7th to 12th days of the disease. CT performance changes with time have a certain regularity, which may indicate the progress and recovery of the disease. 90% of patients still observed residual lung abnormalities in CT images at the time of discharge. There were 4 confirmed cases where the CT images did not show the lesion; hence, CT cannot be used as a basis for judging COVID-19 as a single tool.

Effects of natural capsicum extract on growth performance, nutrient utilization, antioxidant status, immune function, and meat quality in broilers.

This research was conducted to determine the effects of natural capsaicin extract (NCE) as an alternative to the antibiotic (chlortetracycline, CTC) on growth performance, antioxidant capacity, immune function, and meat quality of broiler chickens. A total of 168 one-day-old Arbor Acre male broiler chickens with an average weight of 46.4 ± 0.6 g were randomly allotted to 3 dietary treatments, with 7 replicates per treatment and 8 broilers per pen. These 3 dietary treatments included a corn-soybean meal basal diet (CON), a basal diet + 75 mg/kg CTC (CTC), and a basal diet + 80 mg/kg NCE (NCE). Broilers from the NCE group showed higher average daily gain compared to broilers from the CON group at all stages (P < 0.05). On d 42, NCE supplementation improved dietary nitrogen-corrected apparent metabolizable energy compared to nonsupplemented or CTC-supplemented diets (P < 0.05). The digestibility of organic matter and crude protein were higher in the NCE diet than in the CON or CTC diets (P < 0.05). Higher relative weight of bursa of Fabricius was observed in broilers fed NCE diets compared with CON (P < 0.05). Pancreatic trypsin and lipase activities were significantly increased in the NCE group compared with those in the CON group (P < 0.05). The value of lightness (L*) of breast muscles from broilers fed NCE diets was significantly lower compared to those fed CON diets (P < 0.05). Broilers fed NCE diets also had higher levels of serum total antioxidant capacity, glutathione peroxidase, superoxide dismutase, and lower levels of interleukin-1ß, and tumor necrosis factor-α compared with broilers fed CON diets (P < 0.05). The liver catalase activity of broilers was also significantly increased in the NCE group than the CON group (P < 0.05). In addition, broilers from NCE group had lower concentrations of serum urea-N, low-density lipoprotein cholesterol, and total cholesterol, and higher concentration of growth hormone compared with those from the CON group (P < 0.05). Therefore, we concluded that supplementation of 80 mg/kg of NCE in diets could improve growth performance, nutrient digestibility, antioxidant status, immune function, and meat quality in broilers.

Effects of wheat bran in comparison to antibiotics on growth performance, intestinal immunity, barrier function, and microbial composition in broiler chickens.

This experiment was conducted to evaluate the effects of wheat bran (WB) and antibiotics on growth performance, intestinal immunity, barrier function, and microbial composition in broiler chickens. A total of 168 one-day-old male Arbor Acre chicks were allocated to 3 treatments consisting of 7 replicates with 8 birds per replicate. The 3 treatments were: an antibiotic-free control diet (control, CON), CON + 75 mg/kg chlortetracycline as an antibiotic growth promoter (AGP), and CON + 3% WB. Birds fed AGP and WB had greater (P < 0.05) ADG during days 1 to 21 and lower (P < 0.05) feed-to-gain ratio during each phase than those fed CON. The WB supplementation reduced (P < 0.05) serum concentrations of tumor necrosis factor-α and diamine oxidase activity compared with CON on both day 21 and 42. The AGP and WB supplementation decreased (P < 0.05) interleukin-1ß concentration in jejunal mucosa on day 21 and increased (P < 0.05) secretory immunoglobulin A concentration in jejunal mucosa on day 21 and 42. The relative expression of occludin in jejunal mucosa was upregulated (P < 0.05) in WB than in CON on day 21. Moreover, both AGP and WB supplementation upregulated (P < 0.05) the relative expression of zonula occludens-1 in jejunal mucosa on day 21 and 42. The WB supplementation enhanced the α-diversity of cecal microbiota, as evidenced by the increased Shannon index (P < 0.05). At the phylum level, the phylum Firmicutes was enriched (P < 0.05) in WB. At the genus level, the WB supplementation enriched (P < 0.05) Lachnoclostridium and Butyricicoccus. The WB supplementation increased (P < 0.05) cecal total short chain fatty acids concentrations on day 21 and 42, and butyric acid concentrations on day 42 compared with CON. Collectively, supplementation of 3% WB could promote growth by improving intestinal immunity, barrier function, and microbial composition in broilers. Thus, WB may have a role in replacing antibiotics for improved growth performance and intestinal health in broilers.

Effect of chestnut wood extract on performance, meat quality, antioxidant status, immune function, and cholesterol metabolism in broilers.

Plant extracts have been proved as natural antioxidants resources as well as alternative feed additives in livestock and poultry species. Chestnut wood extract (CWE) as a source of hydrolysable tannic acid was used to evaluate the growth performance, nutrient retention, meat quality, antioxidant status, and immune function of broilers. A total of 168, day-old Arbor Acre male broilers (weight 46.59 ± 0.44 g) were randomly divided to 3 treatments, 7 replicate pens per treatment, 8 broilers per pen. The treatments contain a control diet, CON (corn-soybean meal basal diet); an antibiotic diet, CTC (basal diet + 75 mg/kg chlortetracycline); and chestnut wood extract diet, CWE (basal diet + 1,000 mg/kg chestnut tannins). At the finisher phase, final body weight was higher (P < 0.05) in CWE supplemented diet than in CON. Average daily body weight gain was higher (P < 0.05) and feed gain ratio was lower (P < 0.05) in broilers fed CWE than in those fed CON at the finisher phase. Crude protein digestibility was higher (P < 0.05) in broilers offered CWE than that in broilers fed CON and CTC diets. Breast muscle pH value at 24 h (pH24 h) was higher (P < 0.05) in broilers fed CWE than that in those fed CON and CTC diets. The bursa weight was higher (P < 0.05) in broilers offered CWE than that in those fed CON and CTC. Total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) values were higher (P < 0.05) in both breast muscle and thigh muscle of broilers offered CWE supplemented diet than those in broilers fed CON and CTC diets. Similarly, broilers offered with CWE diets showed higher (P < 0.05) T-AOC, GSH-PX, and SOD value in serum than those fed CON and CTC diets. Serum concentration of IgG was higher (P < 0.05) in broilers offered with CWE diets than that in those fed CON and CTC diets. Total cholesterol, low-density lipoprotein cholesterol, and urea-N concentration were lower (P < 0.05) in broilers offered CWE diet than those in broilers fed CON and CTC diets. It was recommended to supply CWE at the 1,000 mg/kg level for improving antioxidant status, cholesterol metabolism, and growth performance without affecting normal meat quality in broilers.

[Analysis of physical examination data of cardiopulmonary function of graphene workers and health management measures].

Objective: To understand the occupational health examination of graphene workers in a certain enterprise and provide evidence for occupational disease prevention and treatment. Methods: In September 2019, the physical examination data of 54 workers exposed to graphene dust in an occupational health examination of a graphene enterprise from January to August 2019 were collected, and the differences of various physical examination indexes of each group were examined by χ(2) test. Results: The company had a total of 54 graphene workers, and 15 cases of occupational health examination abnormalities, and the abnormal rate was 27.8%. 7 of which had abnormal central functions, and 11 cases of abnormal pulmonary function. 3 of them had abnormal heart function and abnormal lung function. With the increase of age and working age, the abnormal rates of cardiac and pulmonary functions were increased (χ(2)=0.042, P<0.05) . Conclusion: With the development and mass production of graphene, the opportunities for workers to expose to graphene are increasing, and the health of workers is also threatened. Therefore, the occupational disease prevention work of staff should be emphasized.

Circular RNA circ_0000615 knockdown suppresses the development of nasopharyngeal cancer through regulating the miR-338-3p/FGF2 axis.

Nasopharyngeal cancer (NPC) is a type of head and neck cancer with a high rate of metastasis. Circular RNAs (circRNAs) were reported to be related to the development of human cancers. This research aimed to investigate the functional mechanism of circRNA circ_0000615 in NPC. The gene expression was examined by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was employed to assess cell proliferation ability. Transwell assay was used to measure cell migratory and invasive abilities. Furthermore, the interaction between miR-338-3p and circ_0000615 or fibroblast growth factor 2 (FGF2) was predicted by starBase v.2.0 and then confirmed by the dual-luciferase reporter assay. Besides, the mouse xenograft experiment was carried out to explore the effect of circ_0000615 on tumor growth in vivo. We detected increased levels of circ_0000615 and FGF2, along with a decreased level of miR-338-3p in NPC tissues and cells. Circ_0000615 knockdown suppressed the proliferation, migration, invasion, and EMT of NPC cells. Interestingly, circ_0000615 interacted with miR-338-3p, and miR-338-3p targeted FGF2. Circ_0000615 inhibited miR-338-3p expression to upregulate the FGF2 level. Furthermore, both miR-338-3p depletion and FGF2 overexpression weakened the effect of circ_0000615 knockdown on NPC cell progression. Besides, circ_0000615 knockdown repressed tumor growth in vivo. In conclusion, our findings demonstrated that circ_0000615 knockdown suppressed the growth of NPC cells via modulating miR-338-3p/FGF2 axis, providing a theoretical basis for the treatment of NPC.

[Clinical analysis of seven cases of H1N1 influenza-associated encephalopathy in children].

Objective: To investigate the clinical manifestations, diagnosis, and treatment of H1N1 influenza A-associated encephalopathy (IAE) in children. Methods: The clinical manifestations, laboratory tests, cranial magnetic resonance imaging (MRI), electroencephalography (EEG) examinations and treatments of seven children with H1N1 IAE hospitalized in Guangzhou Women and Children's Medical Center from December 2018 to January 2019 were retrospectively analyzed. Results: Five of the seven children with H1N1 IAE were female. The age at admission was 4 years and 5 months (range 7 months-9 years). Neurological symptoms occurred simultaneously or early (0-3 days) after the flu-like symptom appeared. The main clinical manifestations of neurological symptoms were seizures (repeated seizures in five cases and status convulsion in two cases, including one case of unexpected fever and repeated seizures in a nine-year old girl) accompanied with altered consciousness (drowsiness in five cases and coma in two cases). Cranial MRI in three cases displayed multifocal lesions, mainly in the bilateral thalamus, brainstem and cerebellar hemisphere. MRI also showed reversible splenial lesion in the corpus callusumin in three cases. EEG tracings were characterized by diffuse slow wave activity in four cases, and status epilepticus was monitored in one case. All the 7 cases were treated with oral oseltamivir. Three cases were treated with pulsed methylprednisolone and intravenous immunoglobulin. One case was treated with intravenous immunoglobulin alone and all the patients received oral oseltamivir. All the patients survived, with three patients had minor neurological sequelae at discharge. Conclusions: The main clinical manifestations of H1N1 IAE are seizures and altered consciousness. Cranial MRI combined with EEG is helpful for early diagnosis. Intravenous immunoglobulin and (or) methylprednisolone should be considered for severe cases.

Correlations of the ZEB1 expression with the incidence and prognosis of non-small cell lung cancer.

OBJECTIVE: In order to investigate the role of the zinc finger E-box-binding homeobox 1 (ZEB1) expression in the incidence of non-small cell lung cancer (NSCLC), the effects of the ZEB1 expression on the pathogenesis and prognosis of NSCLC were investigated. PATIENTS AND METHODS: Correlations of the expression of ZEB1 in 88 clinical patients with NSCLC with clinicopathological features were determined. The patients were followed up for 60 months to study the correlation between the ZEB1 expression and the prognosis of NSCLC patients. To further explore the role of the ZEB1 expression in the incidence of NSCLC, the expressions of ZEB1 and E-cadherin in three NSCLC cell lines (A549, NCI-H1299 and NCI-H1975) and human normal lung epithelial BESA-2B cell line were measured, and the cell invasion ability was detected. The role of ZEB1 in NSCLC cell invasion was verified through the knockdown of ZEB1 by the short hairpin ribonucleic acids (shRNAs). In addition, its effects on the proliferation and apoptosis of NSCLC cells were confirmed by the overexpression of ZEB1. RESULTS: The expression of ZEB1 in NSCLC tissues was remarkably higher than that in normal tissues, and the expression level of ZEB1 was significantly related to the cancer stage and tumor size. The lower the expression of ZEB1 was, the higher the overall survival rate and the longer the survival time would be. The expression of ZEB1 was negatively correlated with that of E-cadherin in cell lines, and ZEB1-shRNAs markedly reduced the invasion ability of NSCLC cells. The overexpression of ZEB1 resulted in an increase in the proliferation activity and a significant decrease in the apoptosis of A549 cells. CONCLUSIONS: The expression of ZEB1 is closely related to the incidence and prognosis of NSCLC. Increased expression of ZEB1 is helpful for promoting the invasion of NSCLC and enhancing the proliferation activity of NSCLC.

How to make an accurate diagnosis of fetal pyriform sinus fistula in utero: experience at a single medical center in mainland China.

OBJECTIVE: The aim of this study was to make an accurate diagnosis of pyriform sinus fistula (PSF) for prenatal diagnosis. STUDY DESIGN: Medical records were reviewed for all 35 pyriform sinus fistula patients presenting between 2011 and 2017. Ultrasonography (US), fetal magnetic resonance imaging (MRI) and karyotyping were offered during gestation, while computer tomography (CT) and barium esophagography were performed after birth. RESULTS: Patients included 21 males (60%) and 14 females (40%) with a sex ratio of 1.5:1. The lesion was located on the left side in 32 (91.4%) cases, the right side in 2 (5.7%), and was bilateral (2.8%) in only one case. The sensitivity of CT, MRI, ultrasonography and barium esophagography were 100% (35/35), 69.2% (9/13), 22.9% (8/35), and 80% (20/25), respectively. If the diagnosis was correct, there was almost no recurrence after treatment. Karyotype analysis of all fetuses was normal. CONCLUSIONS: Pyriform sinus fistula is more commonly seen in the left side. Compared with ultrasonography, MRI has more advantages in prenatal diagnosis, and it is more accurate in postpartum CT examination. The outcome of children with pyriform sinus fistula may be guarded when it correct diagnosis.

[Clinical analysis of paraneoplastic neurological syndrome associated with thymoma].

Objectives: To investigate the clinical features, diagnosis and treatment of antibody mediated paraneoplastic neurological syndrome associated with thymoma. Methods: From 2012 to 2017, the paraneoplastic antibody and neuron antibody were tested from both blood and cerebrospinal fluid (CSF) in consecutive patients clinically suspected with neurological paraneoplastic syndromes/unknown encephalitis in Peking Union Medical College Hospital.The clinical data, lab results, electrophysiological examinations, imaging features, treatment and clinical prognosis were collected.In this study, the patients who met the diagnostic criteria of both thymoma and neurological paraneoplastic syndrome were retrospectively analyzed.The functional severity was assessed by modified Rankin score(mRS). Results: Six patients (4 female and 2 male) were included for the analysis.Of them, 4 patients presented with limbic encephalitis, 1 with peripheral neuropathy accompanying with myasthenia gravis and 1 with spinal and cerebella degeneration.Three patients were in severe condition (mRS=5). Positive α-amino-3-hydroxy-5-methyl-4-iso xazolepropionic acid receptor, antibody (AMPAR) was identified in 3 patients(2 in blood and CSF, 1 in blood), CV2 plus acetylcholine receptor antibody (ACH-Ab) positive in blood was seen in 1 patient and positive N-methyl-D-aspartate receptor (NMDAR) in blood and CSF in 1 patient. Brain MRI showed abnormalities in 2 patients.The CT scan revealed thymoma in all 6 patients.All patients received intravenous immunogloblin (IVIG) and/or glucocorticoid immediately after diagnosis.Furthermore, thymectomy was performed in 5 patients. All the patients were remarkably improved with mRS 0-1 at discharge.Five patients were clinical stable within follow-up of 1-2.5 years.However, one patient who refused thymectomy experienced several clinical recurrencs. Conclusions: Antibodies mediated paraneoplastic neurological syndrome/autoimmune encephalitis can occur in patients with thymoma with typical characteristics. The correlated antibody test and chest CT might be important in patients clinically suspected with neurological paraneoplastic syndrome or encephalitis with unknown reason. Surgical thymectomy combining with active immunological treatment may play a role in the favorable prognosis, even in those clinical severe patients.