[Activation of HIF-1α/ACLY signaling axis promotes progression of clear cell renal cell carcinoma with VHL inactivation mutation].

Objective: To explore the potential pathogenesis of clear cell renal cell carcinoma (ccRCC) based on the HIF-1α/ACLY signaling pathway, as well as to provide new ideas for the treatment of ccRCC. Methods: Seventy-eight ccRCC cases diagnosed at the First Affiliated Hospital of Soochow University, Suzhou, China were collected. The VHL mutation was examined using exon sequencing. The expression of HIF-1α/ACLY in VHL-mutated ccRCC was evaluated using immunohistochemical staining and further validated in VHL-mutated ccRCC cell lines (786-O, A498, UM-RC-2, SNU-333, and Caki-2) using Western blot. The mRNA and protein levels of ACLY were detected using real-time quantitative PCR and Western blot after overexpression or interference with HIF-1α in ccRCC cell lines. HeLa cells were treated with CoCl2 and hypoxia (1%O2) to activate HIF-1α and then subject to the detection of the ACLY mRNA and protein levels. The potential molecular mechanism of HIF-1α-induced ACLY activation was explored through JASPAR database combined with chromatin immunoprecipitation assay (ChIP) and luciferase reporter gene assay. The effect of HIF-1α/ACLY regulation axis on lipid accumulation was detected using BODIPY staining and other cell biological techniques. The expression of ACLY was compared between patients with ccRCC and those with benign lesions, and the feasibility of ACLY as a prognostic indicator for ccRCC was explored through survival analysis. Results: Exon sequencing revealed that 55 (70.5%) of the 78 ccRCC patients harbored a VHL inactivation mutation, and HIF-1α expression was associated with ACLY protein levels. The protein levels of ACLY and HIF-1α in ccRCC cell lines carrying VHL mutation were also correlated to various degrees. Overexpression of HIF-1α in A498 cells increased the mRNA and protein levels of ACLY, and knockdown of HIF-1α in Caki-2 cells inhibited the mRNA and protein levels of ACLY (P<0.001 for all). CoCl2 and hypoxia treatment significantly increased the mRNA and protein levels of ACLY by activating HIF-1α (P<0.001 for all). The quantification of transcriptional activity of luciferase reporter gene and ChIP-qPCR results suggested that HIF-1α could directly bind to ACLY promoter region to transcriptionally activate ACLY expression and increase ACLY protein level (P<0.001 for all). The results of BODIPY staining suggested that the content of free fatty acids in cell lines was associated with the levels of HIF-1α and ACLY. The depletion of HIF-1α could effectively reduce the accumulation of lipid in cells, while the overexpression of ACLY could reverse this process. At the same time, cell function experiments showed that the proliferation rate of ccRCC cells with HIF-1α knockdown was significantly decreased, and overexpression of ACLY could restore proliferation of these tumor cells (P<0.001). Survival analysis further showed that compared with the ccRCC patients with low ACLY expression, the ccRCC patients with high ACLY expression had a poorer prognosis and a shorter median survival (P<0.001). Conclusions: VHL mutation-mediated HIF-1α overexpression in ccRCC promotes lipid synthesis and tumor progression by activating ACLY. Targeting the HIF-1α/ACLY signaling axis may provide a theoretical basis for the clinical diagnosis and treatment of ccRCC.

[Value of autocrine motility factors in the prediction of the disease progression of PBC- associated hepatocellular carcinoma].

Objective: To clarify the value of autocrine motility factor (ATX) in predicting the disease progression of primary biliary cholangitis (PBC)-associated hepatocellular carcinoma (HCC). Methods: A prospective cohort of 179 newly diagnosed autoimmune liver disease (PBC) patients admitted to the Department of Hepatology at the Fifth Medical Center of the People's Liberation Army General Hospital from January 2016 to January 2018 was selected. All PBC patients received ursodeoxycholic acid (UDCA) treatment and were followed up.The endpoint of the follow-up was the occurrence of primary liver cancer. The relationship between ATX and the clinical characteristics of patients and its significance in predicting disease progression and HCC were analyzed. Results: The peripheral blood ATX level was significantly higher in PBC patients than that of alcoholic cirrhosis (t = 3.278, P = 0.001) and healthy controls (t = 6.594, P < 0.001), but there was no significant difference in ATX levels compared with patients with non-PBC- associated HCC (t = -0.240, P = 0.811). The expression of ATX in liver tissue of PBC patients was significantly higher than that of healthy individuals (Z = -3.633, P < 0.001) and patients with alcoholic cirrhosis (Z = -3.283, P < 0.001), while the expression of ATX in the advanced stage was significantly higher than that in early-stage PBC patients (Z = -2.018, P = 0.034). There was a significant difference in baseline ATX levels between PBC patients without HCC and PBC patients with HCC (228.451 ± 124.093 ng/ml vs. 301.583 ± 100.512 ng/ml, t = 2.339, P = 0.021). Multivariate logistic regression analysis showed that ATX was an independent predictor of PBC progression to HCC (OR = 1.245, 95%CI 1.097-1.413). The baseline peripheral blood ATX level in predicting AUROC of PBC-associated HCC was 0.714, 95%CI 0.597-0.857 and the sensitivity and specificity were 84.6%, and 59.0%, respectively. The optimal cutoff value for predicting serum ATX levels in the occurrence of HCC was 235.254 ng/ml. Conclusion: Patients with PBC have significantly higher levels of ATX expression in their peripheral blood and liver tissue, which can be utilized to assess treatment effectiveness and predict disease progression.

Temperature management of intraoperative cardiopulmonary bypass in valve replacement surgery: a retrospective analysis of the impact on postoperative organ function.

OBJECTIVE: This study aimed to systematically analyze the effects of cardiopulmonary bypass (CPB) at different temperatures on the function of different organs in patients after heart valve replacement and to investigate its safety and feasibility. PATIENTS AND METHODS: The data of 275 heart valve replacement surgery patients who underwent static suction compound anesthesia under CPB between February 2018 and October 2019 were retrospectively analyzed and divided into normothermic CPB anesthesia group (group 0), shallow hypothermic CPB anesthesia group (group 1), medium hypothermic CPB anesthesia group (group 2), and deep hypothermic CPB anesthesia group (group 3) according to the different intraoperative CPB temperatures. The basic preoperative conditions, cardiac resuscitation, number of defibrillations, postoperative ICU stay, postoperative hospital stay, and postoperative evaluation of different organ functions, such as heart, lung, and kidney functions, were analyzed and studied in each group. RESULTS: The comparison of preoperative and postoperative pulmonary artery pressure and left ventricular internal diameter (LVD) was statistically significant in each group (p < 0.05), and the postoperative pulmonary function pressure was statistically significant in group 0 compared with groups 1 and 2 (p < 0.05). The preoperative glomerular filtration rate (eGFR) and the eGFR on the first postoperative day were statistically significant in all the groups (p < 0.05), and the eGFR on the first postoperative day in groups 1 and 2 were statistically significant (p < 0.05). CONCLUSIONS: The control of appropriate temperature during CPB was associated with the recovery of organ function in patients after valve replacement. Intravenous compound general anesthesia with superficial hypothermic CPB might be more beneficial in recovering cardiac, pulmonary, and renal functions.

[Expression of interleukin-34 in tongue squamous cell carcinoma and its clinical implications].

OBJECTIVE: To investigate the expression of interleukin- 34 (IL-34) in tongue squamous cell carcinoma (TSCC) and its clinical implications. METHODS: Serum IL-34 level was detected in 36 patients with TSCC and 36 healthy individuals using enzymelinked immunosorbent assay (ELISA). The expressions of IL-34 mRNA and protein levels in TSCC and adjacent tissues were examined in 41 patients using real-time fluorescence quantitative PCR (qRT-PCR) and immunohistochemistry (IHC), and their correlation with the clinicopathological features of the patients was further analyzed. Informatic analysis of the differentially expressed genes related with IL-34 in TSCC was carried out based on String database, LinkedOmics database and GEO database, and GO functional analysis and KEGG signaling pathway enrichment analysis were performed using Webgestalt database. RESULTS: The serum level of IL-34 was significantly lower in TSCC patients than in the healthy individuals (P < 0.001), and its expression level was also significantly lower in the tumor tissues than in the adjacent tissues (P < 0.001). The expression level of IL-34 in TSCC tissues was related with lymph node metastasis and TNM staging (P < 0.05), but not with age, gender, smoking, drinking, or tumor size (P > 0.05). Informatic analysis suggested that IL-34 had the strongest correlation with CSF1R and PTPRJ. IL-34 and its related genes in TSCC were enriched mainly in bone marrow cell differentiation, collagen-containing extracellular matrix, and cytokine binding and signal receptor activator activity. KEGG signaling pathway enrichment showed that IL-34 and the related differentially expressed genes were involved mainly in osteoclast differentiation, protein polysaccharide in cancer, and the MAPK signaling pathway. CONCLUSION: IL-34 is lowly expressed in TSCC and participates in the occurrence and progression of TSCC, and can be potentially used as a new diagnostic biomarker and therapeutic target for TSCC.

Study on the correlation between dietary structure and sleep in patients with insomnia disorder.

OBJECTIVE: Insomnia disorder (ID) is a persistent difficulty sleeping, often accompanied by anxiety and depression, which seriously reduces a person's quality of life. Dietary changes in insomnia patients have been a concern. To explore the rationality of diet in patients with ID and its correlation with insomnia in ID patients. PATIENTS AND METHODS: This study included 216 patients diagnosed with ID and 197 individuals as the healthy control (HC) group who attended the neurology outpatient clinic or sleep clinic at Henan Provincial People's Hospital between September 2018 and November 2019. Through the Pittsburgh Sleep Quality Index (PSQI), Insomnia Severity Index (ISI), Hamilton Anxiety Scale (HAMA), and Hamilton Depression Scale (HAMD), sleep and mental conditions were assessed in the ID and HC groups. The dietary intake structure of both groups was observed using the food frequency table. Meanwhile, the relationship between dietary intake and sleep quality was analyzed based on the logistics regression. RESULTS: Individuals in the ID group had significantly higher age, weight, and body mass index compared to the HC group (p<0.01). Individuals within the ID category demonstrated a heightened daily consumption of carbohydrates, grains, tubers, and legumes relative to the healthy control group. In contrast, the intake levels of vegetables, fruits, and nuts were diminished compared to the HC group, with this difference being statistically significant (p<0.01). A positive correlation was observed between the daily consumption of grains, tubers, and legumes and PSQI scores. Conversely, a negative association was found between daily consumption of vegetables and fruits. CONCLUSIONS: ID patients exhibit an elevated intake of carbohydrates, whereas the consumption of vegetables, fruits, and nuts is deficient in comparison to the healthy cohort, implying that a distorted dietary structure might be a contributing factor to ID onset. Sensible and scientific dietary guidance is of considerable significance in preventing the onset of ID and facilitating its management. However, the derived conclusions warrant further extensive research.

[Preparation and in vitro evaluation of FDM 3D printed theophylline tablets with personalized dosage].

OBJECTIVE: To explore the feasibility of preparing different doses of tablets for personalized treatment by fused deposition modeling (FDM) 3D printing technology, and to evaluate the in vitro quality of the FDM 3D printed tablets. METHODS: Three different sizes of hollow tablets were prepared by fused deposition modeling 3D printing technology with polyvinyl alcohol (PVA) filaments. Theophylline was chosen as the model drug. In the study, 20 mg, 50 mg and 100 mg of theophylline was filled into the cavity of the tablets, respectively. The microscopic morphology of the tablets was observed by scanning electron microscopy (SEM). The weight variation of the tablets was investigated by weighing method. The hardness of the tablets was measured by tablet hardness tester. The contents of the drugs in the tablets were determined by ultraviolet and visible spectrophotometry (UV-Vis), and the dissolution apparatus was used to assay the in vitro drug release of the tablets. RESULTS: The prepared FDM 3D printed tablets were all in good shape without printing defects. And there was no leakage phenomenon. The diameter and thickness of the tablets were consistent with the design. The layers were tightly connected, and the fine structure of the formulation could be clearly observed without printing defects by scanning electron microscopy. The average weight of the three sizes of tablets was (150.5±2.3) mg, (293.6±2.6) mg and (456.2±5.6) mg, respectively. The weight variation of the three sizes of tablets were boss less than 5%, which met the requirements; The hardness of the tablets all exceeded 200 N; The contents of theophylline in the three tablets were 98.0%, 97.2% and 97.9% of the dosage (20 mg, 50 mg and 100 mg), and the relative standard deviation (RSD) was 1.06%, 1.15% and 0.63% respectively; The time for 80% drug released from the three dosage of tablets was within 30 min. CONCLUSION: Three different dosages of theophylline tablets were successfully prepared by FDM 3D printing technology in this study. The exploration may bring beneficial for the preparation of personalized dose preparations. We expect that with the development of 3D printing technology, FDM 3D printed personalized tablets can be used in the clinic as soon as possible to provide personalized treatment for patients.

[Analysis of the personality characteristics in patients with laryngopharyngeal reflux disease].

Objective: To explore the basic psychological and personality characteristics of patients with laryngopharyngeal reflux disease (LPRD). Methods: The Minnesota Multiphasic Personality Inventory (MMPI) was used to investigate the psychological personality characteristics of 187 patients with LPRD (Study Group) and 75 healthy subjects (control group) who were treated in the otolaryngology clinic of Wuhan Central Hospital from January 2018 to December 2019. There were 102 males in the study group, aged from 20 to 71 (45.42±10.67) years and 85 females, aged from 18 to 65 (40.97±11.53) years; There were 41 males in the control group, aged from 18 to 67 (41.08±12.03) years and 34 females, aged from 20 to 65 (42.55±12.78) years. SPSS 24.0 software was used for statistical analysis. Results: The T scores of HS hypochondriac scale, D depression scale, Hy hysteria scale, Ma hypomania scale and MAS explicit anxiety scale in LPRD patients were significantly higher than those in healthy controls (t=6.304, 4.293, 5.750, 2.694, 5.178, all P<0.05). Correlation analysis showed that RSI score was positively correlated with T score of HS hypochondriac scale, Hy hysteria scale, Ma hypomania scale and MAS explicit Anxiety Scale (r=0.621, 0.402, 0.393, 0.581, all P<0.05); RFS score was positively correlated with T score of HS hypochondriac scale, Hy hysteria scale and MAS explicit Anxiety Scale (r=0.315, 0.321, 0.375, P<0.05). Conclusions: Compared with healthy people, LPRD patients have special personality psychological characteristics, which are mainly manifested as excessive attention to themselves, exaggerated illness, depression, sensitivity, somatization of psychological problems, irritability and mania. The severity of both subjective symptoms (RSI) and objective signs (RFS) of LPRD patients are related to their psychological personality characteristics.

[A two-stitch continuous suture method for single-lumen ileostomy].

Objective: To explore the value of a two-stitch continuous suture in single- lumen ileostomy. Methods: This was a retrospective cohort study. Data for 98 patients who underwent single-lumen enterostomy were retrospectively collected between 1 January 2021 and 1 May 2022 at Zhujiang Hospital of Southern Medical University. All patients met the indications for prophylactic single-lumen ileostomy. Those older than 80 years of age, with complex underlying diseases, extremely poor systemic conditions who could not tolerate surgery, poor blood supply at the end of the bowel, and severe edema or severe infection at the end of the bowel were excluded. Among the included patients, patients who underwent surgery before 1 October 2021 underwent ileostomy with interrupted suture (control group, n=60), and patients operated on and after 1 October 2021 routinely underwent two-stitch continuous suture ileostomy (two-stitch stoma group, n=38). Two-stitch continuous suture ileostomy is performed as follows: the first continuous suture is used to suture the intestinal seromuscular layer, peritoneum, posterior sheath, and anterior sheath from deep to superficial layers. The bowel wall is then opened. The second continuous suture is used to suture the full thickness of the bowel and the skin. The differences in postoperative ostomy-related complications and operation time were compared between the groups. Results: There were no significant differences in baseline data between the groups (all, P>0.05). The operative time in the two-stitch stoma group was shorter than that of the control group (16.6±2.2 minutes vs. 25.1±2.4 minutes, respectively; t=-17.874;P<0.001). The incidences of mucocutaneous separation, dermatitis, and stoma rebound in the two-stitch stoma group were lower than those of the control group [5.3% (2/38) vs. 31.7% (19/60), χ2=9.633, P=0.002;5.3% (2/38) vs. 28.3% (17/60), χ2=7.923, P=0.005; and 2.6% (1/38) vs. 18.3% (11/60), P=0.026, respectively], while the incidences of parastomal hernia and stoma prolapse, and the postoperative visual analog scale scores in the two groups were similar (all P>0.05). Conclusion: Compared with traditional single-lumen ileostomy, two-stitch continuous suture ileostomy has the advantages of short operation time, simplicity, esthetic appearance of the stoma, and a significant reduction in the postoperative complications associated with ileostomy.

[microRNA let-7g-3p regulates proliferation, migration, invasion and apoptosis of bladder cancer cells by targeting HMGB2].

OBJECTIVE: To explore the molecular mechanism by which microRNA let-7g-3p regulates biological behaviors of bladder cancer cells. METHODS: The expression levels of let-7g-3p in bladder cancer and adjacent tissues, normal bladder epithelial cells (HUC cells) and bladder cancer cells (T24, 5637 and EJ cells) were detected using qRT- PCR. T24 cells were transfected with let-7g-3p mimic or inhibitor, and the changes in cell proliferation, migration, invasion, and apoptosis were examined. Transcriptome sequencing was carried out in cells overexpressing let-7g-3p, and the results of bioinformatics analysis, double luciferase reporter gene assay, qRT-PCR and Western blotting confirmed that HMGB2 gene was the target gene of let-7g-3p. The expression of HMGB2 was examined in HUC, T24, 5637 and EJ cells, and in cells with HMGB2 knockdown, the effect of let-7g-3p knockdown on the biological behaviors were observed. RESULTS: qRT-qPCR confirmed that let-7g-3p expression was significantly lower in bladder cancer tissues and cells (P < 0.01). Overexpression of let-7g-3p inhibited cell proliferation, migration and invasion, and promoted cell apoptosis, while let-7g-3p knock-down produced the opposite effects. Bioinformatics and transcriptome sequencing results showed that HMGB2 was the key molecule that mediate the effect of let-7g-3p on bladder cancer cells. Luciferase reporter gene assay, qRT-PCR and Western blotting all confirmed that HMGB2 was negatively regulated by let-7g-3p (P < 0.01). Knocking down HMGB2 could partially reverse the effect of let-7g-3p knockdown on the biological behaviors of the bladder cancer cells. CONCLUSION: The microRNA let-7g-3p can inhibit the biological behavior of bladder cancer cells by negatively regulating HMGB2 gene.

[Multicenter retrospective study of 38 cases with fumarate hydratase deficiency uterine leiomyoma].

Objective: To investigate the clinicopathological features of fumarate hydratase (FH) deficiency uterine leiomyoma. Methods: The data of 38 patients with FH deficiency uterine leiomyoma were screened and analyzed. The expressions of FH, S-(2-succino)-cysteine (2SC), desmin, p16, p53, CD10 and cell proliferation associated nuclear antigen (Ki-67) proteins were detected by immunohistochemistry, and their clinicopathological features were analyzed retrospectively. Results: (1) Clinical features: the median age of the patients was (42.5±7.4) years old. Twenty-one cases (55%) of them were myomas found in physical examination, and the median maximum diameter of the tumor was 6.0 cm (range: 5.0-7.5 cm); myomectomy was performed in 23 cases (61%), total hysterectomy with or without bilateral appendages in 15 cases (39%); laparoscopic surgery in 27 cases (71%), open surgery in 11 cases (29%); none of the patients had renal cell carcinoma. (2) Histological features: atypical nuclear cells were distributed locally or diffusely, eosinophilic nucleoli and intranuclear inclusion bodies could be seen, glass like globules could be seen in the cytoplasm, nuclear division was 0-4/10 high power field (HPF), and antler like blood vessels and pulmonary edema-like changes could be seen in the stroma. Among 38 patients with FH deficiency uterine leiomyoma, FH was negative in 37 cases (97%), and positive in 1 case (3%); 2SC, desmin, p16, p53, CD10 and Ki-67 showed focal positive expression in 38 cases (100%), including 35 cases (92%) with Ki-67 index<10% and 3 cases (8%) with Ki-67 index ≥10%. (3) Follow-up: 4 cases (11%) recurred, and there was no death. There were significant differences in age, family history, distribution of atypical nuclei and mitosis number between recurrent group and non-recurrent group (all P<0.05). Conclusions: FH deficiency uterine leiomyoma is a rare tumor, which needs pathological examination,immunohistochemical examination and clinical history. Patients younger than 43 years old, with family history, histologically atypical diffuse nuclear distribution and mitotic number ≥3/10 HPF should be alert to the risk of recurrence.

[Relationship between parental reproductive age and the risk of overweight and obesity in offspring].

Objective: To explore the role of parental reproductive age on the risk of overweight and obesity in offspring. Methods: The participants were derived from physical examination data of students aged 6-18 years in seven provinces in China, and questionnaire survey was used to collect demographic characteristics and lifestyle information of the students and their parents. A total of 41 567 children with complete data were included. According to the restricted cubic spline curve, maternal reproductive age was divided into three categories, 14-23, 24-28, and 29-38 years, and paternal reproductive age was divided into 14-23, 24-30, and 31-42 years. Multivariate logistic regression model was used to analyze the association between parental reproductive age and parental nutritional status and the risk of overweight and obesity in offspring. Results: The mean age of 41 567 children was (10.6±3.2) years, and the mean paternal and maternal age were (27.9±4.4) years and (25.8±4.0) years, respectively. The detection rate of overweight and obesity was 23.4%. After adjusting factors of children diet and behaviors, the OR(95%CI)of offspring overweight and obesity in groups of fathers aged 24-30 years and mothers aged 24-28 years was 1.11 (1.04-1.18) and 1.16 (1.08-1.24), respectively. When none parents were overweight and obese, the difference of obesity risk was not statistically significant. When both parents were overweight and obese, the OR(95%CI)of offspring overweight and obesity in groups of fathers aged 24-30 years and mothers aged 14-28 years old was 1.27 (1.00-1.62) and 1.33 (1.07-1.65) respectively. Conclusion: Parental reproductive age and parental overweight and obesity status may both increase the risk of overweight and obesity in offspring, with a significant interaction effect.

Effects of honeycomb extract on the growth performance, carcass traits, immunity, antioxidant function and intestinal microorganisms of yellow bantam broilers.

Although many studies have already described the physiological effects of bee products, such as honey, propolis, pollen, and royal jelly, on livestock farming, the health benefits of the honeycomb are still not fully understood. The problem of drug residues and bacterial resistance caused by the abuse of antibiotics is becoming increasingly serious. For this reason, a safe, green substitute has to be sought. We conducted a comparative study of honeycomb extract (HE) and an antibiotic on growth performance, carcass traits, immunity, antioxidant function and intestinal microorganisms of yellow bantam broilers. A total of four hundred eighty 21-day-old female yellow bantam broilers were randomly divided into 5 groups of 6 replicates of 16 birds each. The 5 groups were as follows, with birds receiving a basal diet supplemented with 150 ppm (mg/kg) of chlortetracycline (CTE), a basal diet without HE (control group), and a basal diet with 0.1%, 0.15%, or 0.2% HE for 60 days. The results showed that HE addition significantly increased average daily feed intake (ADFI), average daily gain (ADG), decrease feed gain ratio (F/G) from 21 to 80 and 51 to 80 days of age compared to the control group, with all 3 HE addition groups having statistically identical values to the antibiotic group. HE implementation dramatically increased spleen index, serum immunoglobulin A (IgA), immunoglobulin M (IgM,), glutathione peroxide (GSH-Px), superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and total cecum bacteria and Lactobacillus compared to the control group, numerically at the same level as, or even better than, the antibiotic group. HE and CTE both markly reduced serum malondialdehyde (MDA) concentration compared to the control group, with higher concentrations of HE reducing the effect more dramatically than antibiotics. Both HE and CTE significantly raised dressed yield compared to the control group. In summary, HE, as a potential antibiotic alternative, improved growth performance, carcass traits, immune function, serum antioxidant capacity and intestinal microorganisms in yellow bantam broilers. According to the cubic regression analyses, the recommended supplemental dose of HE was calculated to be 0.15 to 0.17% for female yellow bantam broilers between 21 and 80 d of age.

[Extranuclear p53 suppresses autophagy through AMPK/mTOR signaling to promote heat stress-induced vascular endothelial cell damage].

OBJECTIVE: To explore the role of extranuclear p53-mediated autophagy suppression by regulating AMPK/mTOR signaling pathway in heat stress (HS)-induced injury of mouse aortic endothelial cells (MAECs). METHODS: Primary cultures of MAECs were pretreated with compound C (an AMPK inhibitor), rapamycin (a mTOR inhibitor) or pifithrin-α (PFT, a selective p53 inhibitor) for 1 h before exposure to HS (43 ℃) for 2 h. The changes in cell viability at different time points after HS were examined using CCK-8 assay, and the protein expressions of P53, LC3-II, Beclin-1, p62 and the AMPK/mTOR signaling proteins were detected using Western blotting. In the animal experiment, C57 mice were pretreated with compound C, rapamycin or PFT and exposed to a high temperature at 40 ℃ to induce HS. The pathological changes in the aorta of the mice were observed with HE staining, and cell apoptosis was detected using TUNEL staining. RESULTS: In cultured MAECs, the cell viability was significantly reduced (P < 0.05) and the mitochondrial fraction of p53 increased while its cytoplasmic fraction decreased progressively over time following HS. HS significantly lowered the expressions of LC3-II and Beclin-1, increased p62 level, suppressed AMPK phosphorylation, and increased mTOR phosphorylation and the expressions of its downstream proteins at 6 h after the exposure (P < 0.05). Pretreatment with compound C significantly inhibited LC3-II and Beclin- 1 expression, enhanced p62 expression, and aggravated HS-induced cell injury and apoptosis in MAECs; rapamycin treatment produced the opposite effects (P < 0.05). PFT treatment significantly enhanced the viability of MAECs and alleviated HSinduced injury and apoptosis; PFT also significantly promoted activation of AMPK phosphorylation, inhibited mTOR phosphorylation and its downstream proteins (P < 0.05), enhanced the expressions of LC3-II and Beclin 1, and inhibited p62 expression in the MAECs (P < 0.05). In C57 mice, HS resulted in swelling, shedding and apoptosis of aortic vascular endothelial cells. Pretreatment with compound C obviously aggravated HS-induced vascular injury and endothelial cell apoptosis, while pretreatment with either rapamycin or PFT significantly alleviated these injuries. CONCLUSION: Autophagy inhibition mediated by extranuclear p53 via inhibiting AMPK activity and activating mTOR signaling participates in HS-induced injury of MAECs.

Associations between severe pulmonary function and residual CT abnormalities in rehabilitating COVID-19 patients.

OBJECTIVE: Coronavirus disease 2019 (COVID-19) spread around the world in 2020. Abnormal pulmonary function and residual CT abnormalities were observed in COVID-19 patients during recovery. Appropriate rehabilitation training is around the corner. The correlation between spirometric impairment and residual CT abnormality remains largely unknown. PATIENTS AND METHODS: A cross-sectional study conducted on the pulmonary function of 101 convalescent COVID-19 patients before discharge. Multivariate analysis was used to establish a scoring system to evaluate the spirometric abnormality based on residual chest CT. RESULTS: Lung consolidation area >25% and severe-type COVID-19 were two independent risk factors for severe pulmonary dysfunction. Besides, a scoring system was established. People scoring more than 12 points have more chances (17 times) to get severe pulmonary function impairment before discharge. CONCLUSIONS: For the first time, a chest CT characteristics-based grading system was suggested to predict the pulmonary dysfunction of COVID-19 patients during convalescence in this study. This study may provide suggestions for pulmonary rehabilitation.

[Modulatory effect of 2-arachidonoylglycerol on voltage-gated sodium currents in rat caudate nucleus neurons with kainic acid-induced injury].

OBJECTIVE: To investigate the modulatory effect of 2-arachidonoylglycerol (2-AG) on voltage-gated sodium currents(VGSCs) in rat caudate nucleus (CN) neurons with kainic acid (KA)-induced injury and explore the molecular mechanism underlying the neuroprotective effect of 2-AG. METHODS: Primary cultures of CN neurons isolated from neonatal SD rats were treated with KA, 2-AG+KA, RIM (a CB1 receptor antagonist) +2-AG+KA, or vehicle only (as control).After 7 days in primary culture, the neurons were treated with corresponding agents for 12 h (RIM and 2-AG were added at the same time; KA was added 30 min later) before recording of current density changes, current-voltage characteristics, activation and inactivation kinetics of VGSCs (INa) using whole-cell patch clamp technique. RESULTS: In cultured CN neurons, KA significantly increased current density of VGSCs (P=0.009) as compared with vehicle treatment.KA also produced a hyperpolarizing shift in the activation curve of INa and significantly increased the absolute value of V1/2 for activation (P=0.008).Addition of 2-AG in the culture medium obviously prevented KA-induced increase of INa (P=0.009) and hyperpolarizing shift in the activation curve of INa, and significantly reduced the value of V1/2 for activation(P=0.009)in a CB1 receptor-dependent manner.2-AG alone did not affect the density, activation or deactivation of VGSCs in rat CN neurons. CONCLUSION: In excitotoxic events, endogenous 2-AG can offer neuroprotection by modulating VGSCs in the CN neurons through a CB1 receptor-dependent pathway.

[Renal mucinous tubular and spindle cell carcinoma: clinicopathological and whole exome sequencing analyses].

Objective: To explore the clinicopathological characteristics, immunophenotype, diagnosis and differential diagnosis of renal mucinous tubular and spindle cell carcinoma (MTSCC), and to explore the all-exon mutations, microsatellite stability and tumor mutational burden (TMB) in MTSCC cases. Methods: The data of 5 patients with MTSCC that were submitted to the Department of Pathology, First Affiliated Hospital of Soochow University, China from January 2008 to May 2020, were reviewed and analyzed. The whole exome sequencing (WES) was conducted in all patients, while 3 of them were subject to the analyses of microsatellite stability and TMB. Results: Among the 5 patients, 3 were males and 2 were females. They were 37-76 years old. The maximum diameter of the tumor was 3.5-6.0 cm. The borders of the tumors were well defined. Microscopically, MTSCC was characterized by tubular structure, spindle cell and mucinous stroma, and the nuclear grade of tumor cells was overall low. The average follow-up was 15 months, and no recurrence or metastasis was found. Immunohistochemistry showed that all 5 cases were positive for broad-spectrum cytokeratin (CKpan), cytokeratin (CK)7, CK19, vimentin, PAX8, and P504s (varying expression levels), and the Ki-67 positive index was low. The WES of 5 cases showed that NF2 and PTPN14 exhibited higher mutation rates, which were 3/5 and 2/5, respectively. The microsatellite stability analysis indicated that the 3 cases were all microsatellite stable, and the TMB analysis showed that the TMB of the 3 cases were all <9 mut/Mb. Conclusions: MTSCC is a unique, low-grade pleomorphic kidney tumor. The WES analyses suggest that NF2 and PTPN14 have a higher mutation rate, indicating that the occurrence and development of MTSCC may be closely related to the Hippo pathway. The analysis of microsatellite stability indicates that there is no significant relationship between microsatellite stability and MTSCC, and the TMB analysis suggests that MTSCC patients may not benefit from immunotherapy.

[Immune pathogenesis of primary biliary cholangitis].

Primary biliary cholangitis (PBC) is an autoimmune liver disease, mainly characterized by chronic progressive cholestasis. The root cause of PBC is the loss of immune tolerance to autoantigen E2 subunit of pyruvate dehydrogenase (PDC-E2). The unique immunobiological characteristics of intrahepatic bile duct epithelial cells make it an active participant in the pathogenesis of PBC. In recent years, the detection rate of PBC has been increasing year by year, but the clinical situation of ursodeoxycholic acid monotherapy has not changed. Therefore, an in-depth understanding of the immune pathogenesis of PBC will help clinicians better prevent and treat diseases.

[Epidemiological characteristics of infectious diseases of group A, B and C among Chinese students' population].

OBJECTIVE: To analyze the secular trend, ranking change, age- and regional- characteristics of leading infectious diseases among Chinese students population aged 6 to 22 years from 2008 to 2017. METHODS: Data were drawn from the national surveillance from 2008 to 2017, and the participants were students aged from 6 to 22 years who were diagnosed with notifiable infectious diseases. A total of 40 infectious diseases were classified into three groups based on national notifiable infectious diseases classification of A, B and C. The morbidity and mortality rates from infectious diseases were calculated using the numbers of students published by the ministry of education as the denominator. The age- and province-specific infectious diseases with the highest incidence were selected as the leading infectious diseases for analysis. RESULTS: From 2008 to 2017, the incidence rate, the number of cases and the number of deaths of infectious diseases among the boys aged 6-22 years in China were higher than that of the girls, and the overall trend was downward during the study period. The incidence rates in the boys and girls decreased from 2008 to 2015 with decrease of 43.4% and 40.1%, respectively. However, by 2017, the increase rate rebounded with the increases of 47.1% and 53.8%. The rebound trend was mainly caused by the increase of group C of infectious diseases. During the past decade, the top leading three diseases of groups A and B of infectious diseases were viral hepatitis, tuberculosis and dysentery in 2018, respectively, which changed to tuberculosis, scarlet fever and viral hepatitis in 2017. The top leading three infectious diseases in terms of mortality were rabies, tuberculosis, and human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) in 2008, which were transformed into HIV/AIDS, rabies, and tuberculosis in 2017. There was no significantly obvious change in the incidence and mortality order of group C of infectious diseases during the decade. In the analysis of age groups and regions, the leading infectious diseases in groups A and B transferred from viral hepatitis to scarlet fever and tuberculosis, while in group C, mumps and infectious diarrhoea almost always dominated the leading infectious diseases. But in recent years, influenza and hand-foot-and-mouth disease increased significantly in the eastern region. CONCLUSION: During the past decade, China has got remarkable achievements in the prevention and control of children infectious disease. However, the change patterns and characteristics of notifiable infectious diseases among children and adolescents show the urgent need for prevention and control of respiratory tract infectious diseases, HIV/AIDS and other sexually transmitted diseases as well as new emerging infectious diseases in China in the future. This study provides important basis for policy making of Chinese national school-based infectious disease prevention and control mechanism.

Effects of dietary chromium propionate on laying performance, egg quality, serum biochemical parameters and antioxidant status of laying ducks under heat stress.

Recent studies have shown that chromium (Cr) could alleviate the negative effects of heat stress on livestock and poultry, but there is little information available to laying ducks. This study aimed to investigate the effects of dietary addition of chromium propionate on laying performance, egg quality, serum biochemical parameters and antioxidant status of laying ducks under hot (average 32 °C) and humid (average 75% relative humidity) summer conditions. A total of 900 66-week-old weight- and laying-matched Shanma laying ducks were randomly divided into five treatments, each with 6 replicates of 30 individually caged birds. The birds were fed either a basal diet or the basal diet supplemented with either 200, 400, 600, or 800 µg/kg Cr as chromium propionate. All laying ducks were given feed and water ad libitum for 5 weeks. The results showed that dietary supplementation with chromium propionate significantly increased the laying rate and yolk colour score (P < 0.05). Treatment with 400 µg/kg Cr as chromium propionate significantly decreased the feed/egg ratio by 5.4% (P < 0.05). Increased supplemental Cr from 0 to 800 µg/kg resulted in an increase in albumen height and the Haugh unit linearly (P < 0.05). Increased supplemental Cr decreased serum cortisol (P < 0.001, linear; P = 0.008, quadratic), heat shock protein-70 (P < 0.001, linear; P = 0.007, quadratic) and glucose (P = 0.007, linear), whereas it increased serum insulin (P = 0.011, Linear), total protein (P = 0.006, linear; P = 0.048, quadratic) and albumin (P = 0.035, linear; P = 0.088, quadratic). Dietary Cr levels increased the activities of superoxide dismutase and glutathione peroxidase, the total antioxidant capacity linearly and quadratically (P < 0.05). A linear and quadratic (P < 0.05) decrease of the malondialdehyde concentrations in response to dietary Cr level was observed. These results indicated that dietary supplementation of Cr as chromium propionate, particularly at 800 µg/kg could beneficially affect the laying rate, egg quality and antioxidant function, as well as modulate the blood biochemical parameters of laying ducks under heat stress conditions.