Objective To investigate the molecular mechanism through which calenduloside E inhibits hepatocellular carcinoma(HCC)cell proliferation and migration.Methods HCC cell lines HepG2 and Huh7 treated with calenduloside E were examined for changes in cell viability using CCK-8 assay and expressions of GPX4,SLC7A11,LC3,P62 and phosphorylation of Akt/mTOR using Western blotting.The effects LY294002 and Rapamycin(the inhibitor and activator of autophagy,respectively)on proliferation and migration of calenduloside E-treated HCC cells were evaluated using EdU and Transwell assays.The TCGA database was used to explore the expression levels of GPX4 and SLC7A11 in HCC and normal liver tissues and their correlation with the patients'survival outcomes.GPX4 and SLC7A11 expressions were also detected in HCC cells and normal hepatocytes using RT-qPCR and Western blotting.Results Calenduloside E obviously inhibited the viability of HCC cells.GPX4 and SLC7A11 were highly expressed in HCC tissues and cell lines,and their expression levels were negatively correlated with the patients'survival.In HCC cell lines,calenduloside E significantly inhibited the expressions of GPX4 and SLC7A11 proteins,activated the Akt-mTOR pathway,and enhanced the expression of LC3 II.The inhibitory effect of calenduloside E on GPX4 and SLC7A11 expressions was significantly enhanced by rapamycin but attenuated by LY294002.Inhibiting the autophagy pathway obviously diminished the inhibitory effect of calenduloside E on proliferation and migration of HCC cells,while activating this pathway produced the opposite effect.Conclusion Calenduside E inhibits the proliferation and migration of HCC cells by down-regulating GPX4 and SLC7A11 expression via the autophagy pathway.

Objective To investigate the relationship between insomnia,gastrointestinal symptoms,and glycosylated hemoglobin(HbA1c)levels in individuals diagnosed with type 2 diabetes mellitus(T2DM),as well as the related influencing factors.Methods A total of 910 T2DM patients treated in our multicenter from January 2022 to December 2022 were enrolled in this study.General information(gender,age,smoking and drinking history,exercise,course of disease,treatment and complications),HbA1c,Athens Insomnia Scale(AIS)scores and Gastrointestinal Symptoms Rating Scale(GSRS)scores of patients were collected.The differences of sleep and gastrointestinal symptoms between groups were analyzed,and the correlation between the differences and HbA1c was analyzed.Furthermore,the risk factors for non-standard HbA1c were analyzed.Results The AIS score and GSRS score in the HbA1c control group were less than those in the non-standard group(P＜0.01).Insomnia was reported by 37.0％of T2DM patients,and the HbA1c level in the insomnia group was significantly higher than that in the non-insomnia group(10.00％±2.38％vs.8.26％±1.73％,P＜0.01).Gastrointestinal symptoms were present in 57.5％of T2DM patients,and the HbA1c levels in the group with gastrointestinal symptoms were significantly higher than those in the group without gastrointestinal symptoms(9.26％±2.23％vs.8.43％±1.98％,P＜0.01).Furthermore,26.3％of T2DM patients experienced both insomnia and gastrointestinal symptoms.Remarkably,the HbA1c levels in the group with both insomnia and gastrointestinal symptoms were significantly higher than those in the group without either condition(10.18％±2.44％vs.8.45％±1.86％,P＜0.01).Correlation analysis demonstrated a significant association between sleep quality,gastrointestinal function,and HbA1c levels(P＜0.01).The logistic regression analysis result revealed that age,GSRS score,AIS score,and the presence of insomnia combined with gastrointestinal symptoms were independent risk factors for predicting HbA1c≥6.5％(P＜0.01).Having both insomnia and gastrointestinal symptoms concurrently was the strongest risk factor for substandard HbA1c control,and the risk of blood sugar control may increase about 5 times when both appear together.Conclusion Insomnia and gastrointestinal symptoms are common comorbidities in T2DM patients,showing a cross-interfering relationship,and they appear together with poor blood sugar control,interact causally,and amplify each other.

Objective To investigate the molecular mechanism through which calenduloside E inhibits hepatocellular carcinoma(HCC)cell proliferation and migration.Methods HCC cell lines HepG2 and Huh7 treated with calenduloside E were examined for changes in cell viability using CCK-8 assay and expressions of GPX4,SLC7A11,LC3,P62 and phosphorylation of Akt/mTOR using Western blotting.The effects LY294002 and Rapamycin(the inhibitor and activator of autophagy,respectively)on proliferation and migration of calenduloside E-treated HCC cells were evaluated using EdU and Transwell assays.The TCGA database was used to explore the expression levels of GPX4 and SLC7A11 in HCC and normal liver tissues and their correlation with the patients'survival outcomes.GPX4 and SLC7A11 expressions were also detected in HCC cells and normal hepatocytes using RT-qPCR and Western blotting.Results Calenduloside E obviously inhibited the viability of HCC cells.GPX4 and SLC7A11 were highly expressed in HCC tissues and cell lines,and their expression levels were negatively correlated with the patients'survival.In HCC cell lines,calenduloside E significantly inhibited the expressions of GPX4 and SLC7A11 proteins,activated the Akt-mTOR pathway,and enhanced the expression of LC3 II.The inhibitory effect of calenduloside E on GPX4 and SLC7A11 expressions was significantly enhanced by rapamycin but attenuated by LY294002.Inhibiting the autophagy pathway obviously diminished the inhibitory effect of calenduloside E on proliferation and migration of HCC cells,while activating this pathway produced the opposite effect.Conclusion Calenduside E inhibits the proliferation and migration of HCC cells by down-regulating GPX4 and SLC7A11 expression via the autophagy pathway.

Objective:To review the clinical effect of oral mucosa prefabricated urethra in the treatment of complicated hypospadias in adult patients.Methods:A retrospective analysis was performed on adult patients with complicated hypospadias who underwent two-stage surgical repair in the Second Department of Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2011 to December 2020. In the first stage, the urethra was prefabricated with oral mucosa, which was 5.0-6.0 cm in length and 1.0-1.2 cm in width. The second stage was urethral anastomosis, in which scrotal flap was applied to cover the wound. Postoperative follow-up was conducted by telephone or WeChat, including penile appearance and the situation of urination.Results:A total of 114 patients were enrolled, age raging from 18 to 63 years old, with an average age of 28 years. There were 22 cases who had underwent one previous repair operation, 69 cases had two repair operations, and 23 cases had three repair operations. The time from the last operation was 6 months to 60 months, with an average of 25 months. Postoperative complications included urinary fistula in 36 cases, urethral orifice stenosis in 52 cases and penile scar in 58 cases. The scrotal septal skin flap was used in 46 cases and scrotal fascia flap was used in 68 cases. The flaps size ranged from 1.2 cm×3.0 cm to 3.0 cm×5.5 cm. Among the 114 patients, 6 patients were complicated with urinary extravasation, which healed spontaneously in 2-4 weeks; 2 patients developed urinary fistula due to blood supply disorder at the distal end of the scrotal flap, which was repaired half a year later. Other patients healed well. 94 patients were followed up for 6-84 months, with an average of 37 months. All patients had no chordee, smooth urination, no urethral stricture and fistula. All patients were satisfied with the appearance of the penis. Urethral ultrasound and magnetic resonance hydrography were performed in 18 patients. The results showed that the reconstructed urethra was smooth, the mucosa graft had good extension, and there was no residual urine in the urethra.Conclusions:Oral mucosa prefabricated urethra could be used for repair hypospadias in adult patients who failed in repair surgeries, and could obtain good clinical effect.

Objective:To review the clinical effect of oral mucosa prefabricated urethra in the treatment of complicated hypospadias in adult patients.Methods:A retrospective analysis was performed on adult patients with complicated hypospadias who underwent two-stage surgical repair in the Second Department of Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2011 to December 2020. In the first stage, the urethra was prefabricated with oral mucosa, which was 5.0-6.0 cm in length and 1.0-1.2 cm in width. The second stage was urethral anastomosis, in which scrotal flap was applied to cover the wound. Postoperative follow-up was conducted by telephone or WeChat, including penile appearance and the situation of urination.Results:A total of 114 patients were enrolled, age raging from 18 to 63 years old, with an average age of 28 years. There were 22 cases who had underwent one previous repair operation, 69 cases had two repair operations, and 23 cases had three repair operations. The time from the last operation was 6 months to 60 months, with an average of 25 months. Postoperative complications included urinary fistula in 36 cases, urethral orifice stenosis in 52 cases and penile scar in 58 cases. The scrotal septal skin flap was used in 46 cases and scrotal fascia flap was used in 68 cases. The flaps size ranged from 1.2 cm×3.0 cm to 3.0 cm×5.5 cm. Among the 114 patients, 6 patients were complicated with urinary extravasation, which healed spontaneously in 2-4 weeks; 2 patients developed urinary fistula due to blood supply disorder at the distal end of the scrotal flap, which was repaired half a year later. Other patients healed well. 94 patients were followed up for 6-84 months, with an average of 37 months. All patients had no chordee, smooth urination, no urethral stricture and fistula. All patients were satisfied with the appearance of the penis. Urethral ultrasound and magnetic resonance hydrography were performed in 18 patients. The results showed that the reconstructed urethra was smooth, the mucosa graft had good extension, and there was no residual urine in the urethra.Conclusions:Oral mucosa prefabricated urethra could be used for repair hypospadias in adult patients who failed in repair surgeries, and could obtain good clinical effect.

Objective:To review the clinical effect of oral mucosa prefabricated urethra in the treatment of complicated hypospadias in adult patients.Methods:A retrospective analysis was performed on adult patients with complicated hypospadias who underwent two-stage surgical repair in the Second Department of Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2011 to December 2020. In the first stage, the urethra was prefabricated with oral mucosa, which was 5.0-6.0 cm in length and 1.0-1.2 cm in width. The second stage was urethral anastomosis, in which scrotal flap was applied to cover the wound. Postoperative follow-up was conducted by telephone or WeChat, including penile appearance and the situation of urination.Results:A total of 114 patients were enrolled, age raging from 18 to 63 years old, with an average age of 28 years. There were 22 cases who had underwent one previous repair operation, 69 cases had two repair operations, and 23 cases had three repair operations. The time from the last operation was 6 months to 60 months, with an average of 25 months. Postoperative complications included urinary fistula in 36 cases, urethral orifice stenosis in 52 cases and penile scar in 58 cases. The scrotal septal skin flap was used in 46 cases and scrotal fascia flap was used in 68 cases. The flaps size ranged from 1.2 cm×3.0 cm to 3.0 cm×5.5 cm. Among the 114 patients, 6 patients were complicated with urinary extravasation, which healed spontaneously in 2-4 weeks; 2 patients developed urinary fistula due to blood supply disorder at the distal end of the scrotal flap, which was repaired half a year later. Other patients healed well. 94 patients were followed up for 6-84 months, with an average of 37 months. All patients had no chordee, smooth urination, no urethral stricture and fistula. All patients were satisfied with the appearance of the penis. Urethral ultrasound and magnetic resonance hydrography were performed in 18 patients. The results showed that the reconstructed urethra was smooth, the mucosa graft had good extension, and there was no residual urine in the urethra.Conclusions:Oral mucosa prefabricated urethra could be used for repair hypospadias in adult patients who failed in repair surgeries, and could obtain good clinical effect.

Objective:To review the clinical effect of oral mucosa prefabricated urethra in the treatment of complicated hypospadias in adult patients.Methods:A retrospective analysis was performed on adult patients with complicated hypospadias who underwent two-stage surgical repair in the Second Department of Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2011 to December 2020. In the first stage, the urethra was prefabricated with oral mucosa, which was 5.0-6.0 cm in length and 1.0-1.2 cm in width. The second stage was urethral anastomosis, in which scrotal flap was applied to cover the wound. Postoperative follow-up was conducted by telephone or WeChat, including penile appearance and the situation of urination.Results:A total of 114 patients were enrolled, age raging from 18 to 63 years old, with an average age of 28 years. There were 22 cases who had underwent one previous repair operation, 69 cases had two repair operations, and 23 cases had three repair operations. The time from the last operation was 6 months to 60 months, with an average of 25 months. Postoperative complications included urinary fistula in 36 cases, urethral orifice stenosis in 52 cases and penile scar in 58 cases. The scrotal septal skin flap was used in 46 cases and scrotal fascia flap was used in 68 cases. The flaps size ranged from 1.2 cm×3.0 cm to 3.0 cm×5.5 cm. Among the 114 patients, 6 patients were complicated with urinary extravasation, which healed spontaneously in 2-4 weeks; 2 patients developed urinary fistula due to blood supply disorder at the distal end of the scrotal flap, which was repaired half a year later. Other patients healed well. 94 patients were followed up for 6-84 months, with an average of 37 months. All patients had no chordee, smooth urination, no urethral stricture and fistula. All patients were satisfied with the appearance of the penis. Urethral ultrasound and magnetic resonance hydrography were performed in 18 patients. The results showed that the reconstructed urethra was smooth, the mucosa graft had good extension, and there was no residual urine in the urethra.Conclusions:Oral mucosa prefabricated urethra could be used for repair hypospadias in adult patients who failed in repair surgeries, and could obtain good clinical effect.

Objective To investigate the relationship of the expression of peripheral T helper type 9 (Th9) cells and their related cytokines with the severity and clinical course of alopecia areata,and to explore their significance in the occurrence of alopecia areata.Methods From May to December in 2017,74 outpatients with alopecia areata enrolled from Department of Dermatology,Huashan Hospital,Fudan University served as the alopecia areata group,and 57 health checkup examinees in Huashan Hospital served as the control group.Peripheral blood samples were obtained from the patients and controls.Enzyme-linked immunosorbent assay (ELISA) was performed to detect the serum levels of interleukin (IL)-9,IL-4,transforming growth factor (TGF)-β1 and interferon (IFN)-γ,flow cytometry to determine the proportion of Th9 cells (CD4+IL-9+ T helper cells) in peripheral blood mononuclear cells (PBMC),and real-time fluorescence-based quantitative PCR to measure the mRNA expression of IL-9 and PU.1 in the PBMCs.Data were recorded in Microsoft Excel 2017 software,and statistically analyzed with SPSS17.0 software using two-sample t test and Spearman rank correlation analysis.Statistical charts were drawn with Graphpad prism 6 software.Results Compared with the control group,the alopecia areata group showed significantly decreased serum level of IL-9 (190.40 ± 12.33 ng/L vs.288.10 ± 17.38 ng/L,t =4.71,P ＜ 0.01),but significantly increased serum levels of TGF-β1 (6 191.00 ± 355.50 ng/L vs.4 026.00 ± 258.00 ng/L,t =4.41,P ＜ 0.05) and IFN-γ(15.71 ± 3.00 ng/L vs.8.79 ± 0.60 ng/L,t =2.001,P ＜ 0.05).However,there was no significant difference in the serum level of IL-4 between the alopecia areata group and control group (P ＞ 0.05).The serum level of IFN-γ was significantly lower in the patients with severe alopecia areata than in the patients with mild alopecia areata (P =0.02),and the serum level of IFN-γ in the patients with alopecia areata was negatively correlated with the severity of alopecia tool (SALT) score (ru =-0.298,P =0.010).There were no significant differences in the serum levels of IL-9,IL-4 and TGF-β1 between the patients with severe alopecia areata and those with mild alopecia areata (all P ＞ 0.05).The serum levels of IL-9,IL-4,TGF-β1 and IFN-γdid not differ between the patients with active alopecia areata and those with stable alopecia areata,as well as between the patients with clinical course of ＜ 6 months and those with clinical course of ＞ 6 months (P ＞ 0.05).The alopecia areata group showed significantly decreased proportion of Th9 cells in the PBMCs (t =2.04,P =0.045) and mRNA expression of IL-9 and PU.1 (t =2.12,2.178,both P ＜ 0.05) compared with the control group.Condusion The serum level of IL-9 and proportion of peripheral blood Th9 cells both decrease in patients with alopecia areata,and Th9 cells and their related cytokines may be involved in the occurrence of alopecia areata.

The ZNF804A variant rs1344706 has consistently been associated with schizophrenia and plays a role in hippocampal-prefrontal functional connectivity during working memory. Whether the effect exists in the resting state and in patients with schizophrenia remains unclear. In this study, we investigated the ZNF804A polymorphism at rs1344706 in 92 schizophrenic patients and 99 healthy controls of Han Chinese descent, and used resting-state functional magnetic resonance imaging to explore the functional connectivity in the participants. We found a significant main effect of genotype on the resting-state functional connectivity (RSFC) between the hippocampus and the dorsolateral prefrontal cortex (DLPFC) in both schizophrenic patients and healthy controls. The homozygous ZNF804A rs1344706 genotype (AA) conferred a high risk of schizophrenia, and also exhibited significantly decreased resting functional coupling between the left hippocampus and right DLPFC (F(2,165) = 13.43, P < 0.001). The RSFC strength was also correlated with cognitive performance and the severity of psychosis in schizophrenia. The current findings identified the neural impact of the ZNF804A rs1344706 on hippocampal-prefrontal RSFC associated with schizophrenia.
Adult ; Analysis of Variance ; Female ; Functional Laterality ; genetics ; Genotype ; Hippocampus ; diagnostic imaging ; Humans ; Image Processing, Computer-Assisted ; Kruppel-Like Transcription Factors ; genetics ; Magnetic Resonance Imaging ; Male ; Neural Pathways ; diagnostic imaging ; Neuropsychological Tests ; Oxygen ; blood ; Polymorphism, Single Nucleotide ; genetics ; Prefrontal Cortex ; diagnostic imaging ; Psychiatric Status Rating Scales ; Schizophrenia ; diagnostic imaging ; genetics ; physiopathology ; Severity of Illness Index ; Young Adult

Objective:To explore the traits of functional connectivity of resting-state networks in patients of schizophrenia with auditory verbal hallucinations (AVH) by using independent component analysis (ICA).Methods:All patients were met the Diagnostic and Statistical Mannal of Mental Disorders,Fourth Edition,Text Revision (DSM-IV-TR) diagnostic criteria for schizophrenia.Thirty schizophrenia patients with frequent AVH (AVH),24 schizophrenia patients without AVH (non-AVH) and 60 healthy controls (HC) matching with age and gender were analyzed by resting functional magnetic resonance imaging.The AVH were assessed by using Hoffman auditory hallucination scale.By using ICA,auditory network (AUN),language network (LAN),salience network (SAN),right front-parietal network (RFP) and default mode network (DMN) were selected as interesting networks for further analyses.Covariance analysis was used to compare the activity and functional connectivity of multiple resting state networks between three groups,and correlation analysis was used to evaluate the relations between abnormalities of brain and the severity of hallucination.Results:Compared to non-AVH,AVH group showed increased activity in left superior temporal gyrus (STG) and left postcentral gyrus (LPG) in the AUN,and showed decreased activity in right anterior cingulate cortex (ACC) in the SAN (P ＜ 0.05,FDR corrected).The severity of AVH group was associated with activity of the left STG (r =0.43,P ＜0.05) and the right ACC (r =-0.48,P ＜0.01).Furthermore,compared to non-AVH group,there were decreased connectivity between LAN and AUN [(-0.29 ±0.21) vs.(-0.16 ± 0.17),P ＜ 0.05,uncorrected],decreased connectivity between SAN and LAN [(-0.09 ± 0.22) vs.(-0.06 ± 0.18),P ＜ 0.05,uncorrected],increased connectivity between SAN and AUN [(0.30 ± 0.18) vs.(0.15 ± 0.24),P ＜ 0.05,uncorrected] in AVH group.Conclusion:There may be aberrant functional connectivities of AUN,LAN and SAN in schizophrenia patients with frequent AVH,and the occurrence of AVH seems to be related to brain areas involved in language production,speech perception and self-monitoring.