ObjectiveTo investigate the etiology and clinical features of intrahepatic cholestasis and the diagnostic value of whole exome sequencing （WES） through a retrospective analysis of the medical history， pathological results， and gene sequencing data of 62 patients with unexplained intrahepatic cholestasis. MethodsA retrospective analysis was performed for the clinical data of 480 patients who underwent WES due to unexplained liver function abnormalities in Nanjing Second Hospital from January 2017 to December 2023， among whom 62 patients with unexplained intrahepatic cholestasis were selected based on laboratory data， and a confirmed diagnosis was made based on imaging data， pathological findings， and gene sequencing data. The patients with unexplained intrahepatic cholestasis were analyzed in terms of demographic features， clinical manifestation， etiology spectrum， and genetic profile. ResultsA total of 62 patients with unexplained intrahepatic cholestasis were included， among whom there were 35 male patients and 27 female patients， with a median age of 42 （7 — ‍77） years. WES was used to make a definite diagnosis in 21 patients （33.87%）， among whom the patients with familial intrahepatic cholestasis accounted for the highest proportion of 52.38% （11/21）； genetic metabolic disorders were excluded by WES in 34 patients， with drug-induced liver injury and sepsis-associated liver injury accounting for the highest proportion of 55.88% （19/34）， followed by primary biliary cholangitis and primary sclerosing cholangitis accounting for 20.59% （7/34） and intrahepatic bile duct stones accounting for 17.65% （6/34）， while the patients with a lack of confirmed diagnosis accounted for 11.29% （7/62）. A total of 21 novel mutation sites which were not reported in previous articles were identified in this study. ConclusionGenetic metabolic disorders constitute a significant proportion of unexplained intrahepatic cholestasis， and WES plays a crucial role in the diagnosis of unexplained intrahepatic cholestasis.

Objective To investigate the role of PPARγ/GluT4 axis in insulin resistance(IR),cell proliferation and apoptosis of granulosa cells.Methods A total of 45 married women with PCOS who received routine IVF-ET assisted pregnancy treatment in Center of Reproductive Medicine,Qinghai Provincial People's Hospital were enrolled in this study from August 2018 to August 2020.All the patients were divided into IR group(PCOS-IR group,HOMA-IR≥2.57,n=23)and non-IR group(PCOS-NIR group,HOMA-IR<2.57,n=22)according to HOMA-IR.Meanwhile,21 married patients with infertility due to male or fallopian tube factors were enrolled as control group(Con).miR-27a mimics,miR-27a inhibitors(miR-27a inhibitor)and corresponding controls(mimics NC and inhibitor NC)transfected PCOS-IR granulosa cells,which were then divided into miR-27a mimics group,miR-27a inhibitor group,mimics-NC group and inhibitor-NC group.Double luciferase report test confirmed that miR-27a binded to PPARγ.Cell proliferation and apoptosis were detected by CCK-8 method and Annexinv-FITC/PI method.The expression of miR-27a,GluT4,PPARγ,Bax related to B lymphomas-2,Cleaved caspase-3 and B lymphomas-2(Bcl-2)were detected by RT-qPCR and Western blot respectively.Results Compared with Con group,the expression of miR-27a increased(P<0.01),while the expression of PPARγ mRNA and protein decreased in PCOS-NIR and PCOS-IR groups(P<0.01).Compared with PCOS-NIR group,the expression of miR-27a increased(P<0.05),while the expression of PPARγ mRNA and protein decreased in PCOS-IR group(P<0.01).The double luciferase report showed that there was a targeted binding site between PPARγ and miR-27a.Compared with inhibitor-NC group,the cell activity increased at 24 h,48 h,72 h and 96 h in miR-27a inhibitor group(P<0.05 or P<0.01),while the apoptosis rate decreased inmiR-27a inhibitor and mimics-NC group(P<0.05 or P<0.01).Compared with miR-27a inhibitor group,the apoptosis rate increased,and the cell activity decreased at 24,48,72 and 96 h in mimics-NC and miR-27a mimics groups(P<0.05 or P<0.01).Compared with the inhibitor-NC group,the expression of miR-27a,Bax and Cleaved caspase-3 increased(P<0.05 or P<0.01),while the expression of GluT4,PPARγ and Bcl-2 decreased in miR-27a mimics group(P<0.01).In miR-27a inhibitor group,the protein expressions of GluT4,PPARγ and Bcl-2 increased(P<0.05 or P<0.01),while miR-27a,Bax and Cleaved caspase-3 decreased(P<0.01).Compared with miR-27a inhibitor group,the expressions of miR-27a,Bax and Cleaved caspase-3 increased(P<0.01),while the expressions of GluT4,PPARγ and Bcl-2 decreased in mimics-NC and miR-27a mimics groups(P<0.05 or P<0.01).Conclusion The expression level of miRNA-27a is related to IR,cell proliferation,and apoptosis of granulosa cells,which may be related to PPARγ signal path.

Morita therapy has been bom for more than 100 years.Inpatient Morita therapy is highly oper-able and easy to master.It can improve many refractory neuroses through four-stage treatment.But more neuroses are treated in outpatient clinics,and Morita therapy cannot be used in hospitalized patients.Therefore,the formula-tion of expert opinions on outpatient operations is particularly important.This paper is based on domestic and for-eign references,and after many discussions by domestic Morita therapy experts,and then drew up the first version of the expert opinions on operation of outpatient Morita therapy.Meanwhile the operation rule of Morita therapy in three stages of outpatient treatment was formulated:in the etiological analysis stage,under the theoretical guidance of Morita therapy,analyze the pathogenic factors,to improve treatment compliance and reduce resistance;during the operating stage,guide patients to engage in constructive and meaningful actions,realizing the achievement of letting nature take its course principle;in the cultivating character and enriching life stage,pay attention to positive infor-mation,expanding the scope and content of actions,improving the ability to adapt to complex life,and preventing recurrence caused by insufficient abilities.It will lay a foundation for the promotion of Morita therapy in domestic outpatient clinics,so that more patients with neurosis and other psychological diseases could receive characteristic Morita therapy treatment in outpatient clinics.

BACKGROUND:In recent years,High-definition transcranial direct current stimulation(HD-tDCS)has garnered significant attention due to its potential non-invasive modulation of brain function.However,there is still a lack of visual analysis in the literature regarding this technique. OBJECTIVE:To perform a visual analysis of HD-tDCS-related research so as to explore the current status and trends in this field. METHODS:English literature related to HD-tDCS was retrieved from the Web of Science Core Collection database covering the period from January 1,2010 to May 6,2023.The VOSviewer software was used to visualize and analyze the source journals,countries/regions,authors,institutions,cited references,and keywords of the included literature,creating a knowledge map to uncover the research landscape and hotspots. RESULTS AND CONCLUSION:A total of 336 articles were included,showing a consistent increase in the annual publication count of HD-tDCS research.Among these,the United States contributed the highest number of publications(141 articles)with 4 221 citations,followed by China with 70 articles and 401 citations.Brain Stimulation was the most prolific journal(28 articles),Marom Bikson was the most productive author(37 articles),and The City College of New York was the most active institution(35 articles).The most frequently mentioned keywords in the field included motor cortex,regulation,working memory,excitability,and dorsolateral prefrontal cortex.Notable recent keywords in the last 5 years include attention-deficit/hyperactivity disorder,brain networks,and stimulation intensity.Currently,the volume of HD-tDCS research is relatively limited,but is on an upward trajectory,indicating substantial research potential.It is expected that future studies in this domain will continue to focus on the application of HD-tDCS in cognitive and neuropsychiatric disorders,while also exploring its therapeutic mechanisms targeting the motor cortex and dorsolateral prefrontal cortex based on brain network analysis.

BACKGROUND:The abnormal gait of stroke patients seriously affects their propulsive force during walking,which subsequently reduces their walking speed,walking distance,and stability,increases their risk of falls,and seriously affects their quality of life. OBJECTIVE:To review the relevant research on propulsive force deficits in stroke patients with hemiplegia,to summarize the understanding of existing researchers on propulsive force deficits,to analyze the relationship between propulsive force and gait,and finally to explain and compare the latest rehabilitation technologies used to improve propulsive force deficits,providing reference for clinical treatment. METHODS:Relevant literature was retrieved from WanFang,CNKI,PubMed,and Web of Science Core Collection through computer search.The Chinese and English search terms were"propulsive force OR propulsive,stroke OR cerebral infarction OR hemiplegia,walk* OR gait."The search time limit was from 2003 to 2023,and 71 articles were finally included for review and analysis. RESULTS AND CONCLUSION:Training targeting the hip and ankle joints may be more effective for patients'walking function,especially training with the application of flexible exoskeleton robots,but more sufficient evidence is still needed to use propulsion as a prognostic indicator of walking function in stroke patients.Biomechanical variables related to propulsive force include:the hip joint extension angle at terminal stance,ankle joint dorsiflexion torque,and knee joint extension.Damage to the corticospinal tract,cerebellar-cortical pathways,and the reticulospinal tract in hemiplegic patients are associated with reduced propulsive force and gait asymmetry.Propulsive force is crucial for the stability of healthy gait,and a decrease in propulsive force is unfavorable for gait stability.Gait symmetry is correlated with propulsive force,stride length symmetry,trunk displacement,and lower limb swing ability,with propulsive force being a key factor.Propulsive force can serve as a quantitative indicator for assessing the gait of hemiplegic patients,and evaluation of gait using propulsive force is beneficial for the long-term development of walking ability.Main rehabilitation techniques for improving propulsive force include:lower limb exoskeleton robot walking training,treadmill training combined with functional electrical stimulation,adaptive speed treadmill training,biofeedback technology,and whole-body vibration training.Among them,whole-body vibration training and biofeedback technology are more effective.The specific contributions and mechanisms of the hip,knee,and ankle joints in improving propulsive force are still controversial,but it is expected that the contributions of the hip and ankle joints are greater.Focusing on the improvement of propulsive force as a rehabilitation goal may yield more sustainable advancements in walking function.However,several current challenges persist in this field:understanding the neurobiological basis of propulsive force deficits in stroke patients,assessing the long-term efficacy of current rehabilitation techniques for enhancing propulsive force,and determining the most suitable patient populations for the application of major rehabilitation techniques aiming at improving propulsive force.These areas require further exploration by subsequent researchers.

Objective:To explore the effects of type D personality on mild cognitive dysfunction (MCI) in patients with coronary heart disease and the multiple mediating roles of dietary inflammation index (DII) and systemic immune inflammation index (SII) between type D personality and MCI.Methods:A cross-sectional study was used. A total of 321 patients diagnosed with coronary heart disease by coronary angiography in the Second Affiliated Hospital of Harbin Medical University from May 2022 to March 2023 were selected as the study objects by convenient sampling method. General Information Questionnaire, Type-D Personality Scale-14, Montreal Cognitive Assessment, and Semi-Quantitative Food Frequency Questionnaire were employed for data collection. DII and SII were used to evaluate dietary inflammatory potential and systemic inflammatory status, respectively. Structural equation modeling was utilized to analyze the mediating effectsamong type D personality, DII, SII and MCI.Results:A total of 306 valid questionnaires were collected. Among the 306 patients, there were 215 males (70.3%) and 91 females (29.7%), aged 27-70 (59.24 ± 9.16) years old. The structural equation model showed that type D personality could directly influence MCI (effect size = - 1.098, 95% CI - 1.869 - - 0.327), and could also mediate the occurrence of MCI in coronary heart disease patients through the single mediation of DII (effect size = - 0.374, 95% CI - 0.644 - - 0.128) and SII (effect size = - 0.450, 95% CI - 0.806 - - 0.132), as well as the chain mediation of DII and SII (effect size = - 0.146, 95% CI - 0.293 - - 0.027). Conclusions:Type D personality, DII and SII can affect the occurrence of MCI in CHD patients, and DII and SII play multiple mediating roles between type D personality and MCI in patients with coronary heart disease. Clinical and community nurses can improve the unhealthy dietary behavior, reduce the level of inflammation and delay the occurrence and development of MCI by early screening of type D personality in patients with coronary heart disease.

Objective To investigate the effect of agomelatine on the clinical efficacy in elderly patients with acute cerebral infarction(ACI)and comorbid anxiety-depression disorders by regu-lating serum neurotransmitters and nerve cytokines.Methods A total of 160 elderly ACI patients with anxiety and depression symptoms admitted in Pingxiang Second People's Hospital from June 2020 to December 2023 were enrolled in this study.All of them received thrombolysis or interven-tional therapy,and then were randomly divided into control and observation groups,with 80 patients in each group.The control group received conventional psychological intervention,while the observation group was given additional oral administration of agomelatine.Cognitive function,neurological function,neurotransmitters and neuronal cytokines,anxiety and depression scores,sleep quality,quality of life,daily activity ability and adverse reactions were compared between the two groups.Results After intervention,Mini Mental State Examination(MMSE)scores,levels of neuropeptide Y(NPY),5-hydroxytryptamine(5-HT),norepinephrine(NE),dopamine and brain-derived neurotrophic factor(BDNF),36-item Brief Health Questionnaire(SF-36)score,and Bar-thel index scale score were significantly higher in both 2 groups when compared with above indicators before the intervention(P＜0.01).And the MMSE score,NPY,5-HT,NE,dopamine and BDNF levels,SF-36 score and Barthel index scale score were obviously higher in the observa-tion group than the control group(P＜0.01).Both groups obtained notably lower NIHSS score,S100 calcium binding protein B(S100B)and myelin basic protein(MBP)levels,Hamilton Anxiety Rating Scale(HAMA)score,and Hamilton Depression Rating Scale 17(H AMD-17)score and Pittsburgh Sleep Quality Index(PSQI)score after intervention(P＜0.01).And the NIHSS score,S100B and MBP levels,HAMA score,HAMD-17 score,and PSQI score were statistically lower in the observation group than the control group(P＜0.01).During the treatment process,no signifi-cant difference was observed in the incidence of total adverse reactions between the two groups(3.75％vs 6.25％,x2=0.526,P=0.468).Conclusion When agomelatine tablets are indicated for ACI patients with concomitant anxiety-depression disorders,they can effectively rehabilitate cog-nitive function,enhance neurological function,improve sleep quality and quality of life,optimize activities of daily living,eliminate negative emotions,and correct the expression of neurotransmit-ters and neurotrophic factors.

Objective:To investigate the effect of methyltransferase like 14 (METTL14) on the proliferation and invasion of breast cancer (BC) cells by regulating cyclin L2 (Cyclin L2, CCNL2) through m6A modification.Methods:Cancer tissues and paracancerous tissues of BC patients in Yantaishan hospital were collected from Aug. 2018 to Feb. 2020. The expression levels of m6A, METTL14 and CCNL2 in tissues were detected by high performance liquid chromatography/mass spectrometry (HPLC/MS) and qRT-PCR. Dual-luciferase reporter assay, qRT-PCR, and western blot were used to verify the regulatory relationship between METTL14 and CCNL2. RIP experiments verified the regulatory relationship between YTH domain-containing family protein (YTHDF2) and CCNL2. Cell viability was detected by MTT method, and cell invasion ability was detected by Transwell.Results:Compared with normal cells (0.24±0.02) and tissues (0.18±0.02) , BC cells MCF-10A (0.47±0.03, t=11.05, P<0.001) and HS-578T (0.41±0.03, t=8.17, P=0.001) and BC tissues (0.39±0.02, t=12.86, P<0.001) m6A level increased. Compared with normal tissues (1.00±0.26) (0.84±0.07) , METTL14 mRNA (1.57±0.28, t=13.50, P<0.001) and protein levels (1.66±0.11, t=10.89, P<0.001) in BC tissues were significantly increased high. Compared with the control group (100.00±10.11) (1.00±0.12) , the BC cell invasion ability (54.15±6.21, t=6.69, P=0.003) and activity (0.64±0.06, t=4.65, P=0.010) were weakened. Compared with the control group (100±11.05) (1±0.13) , the BC cell invasion ability (175.31±13.45, t=7.49, P=0.002) and activity (2.16±0.16, t=9.75, P=0.002) in the METTL14 overexpression group were enhanced, and the effects of METTL14 on cell invasion (137.41±12.64, t=3.56, P=0.024) and activity (1.64±0.15, t=5.59, P=0.005) were partially reversed after m6A inhibitor treatment change. Compared with normal tissues, CCNL2 expression was down-regulated in BC tissues, and the interaction between CCNL2 and METTL14 was confirmed. Compared with the control group (1.00±0.1) (0.64±0.05) , knockdown of METTL14 could make CCNL2 mRNA (1.67±0.05) . 0.13, t=7.08, P=0.002) and protein (1.09±0.09, t=7.57, P=0.002) were up-regulated. METTL14 knockout enhanced the stability of CCNL2 mRNA through a YTHDF2-dependent pathway, compared with sh-METTL14 group (50.47±5.16) (0.52±0.05) , BC cell invasion ability of sh-METTL14+sh-CCNL2 group (71.69±6.41, t=4.47, P=0.011) and activity (0.64±0.05, t=2.94, P=0.042) were improved. Conclusion:METTL14 inhibits the expression of CCNL2 through m6A modification to enhance the invasion and activity of BC cells.

Objective:To preliminarily investigate the effect of circIGF2BP3 on autophagy in photoaged dermal fibroblasts.Methods:Human dermal fibroblasts were isolated from circumcised foreskin tissues from 6 children in the Department of Urological Surgery, Third Affiliated Hospital of Sun Yat-sen University. An ultraviolet A (UVA) -induced photoaged human dermal fibroblast model (UVA radiation group) was established by repeated UVA radiation at a dose of 10 J/cm 2 for 14 consecutive days, and human dermal fibroblasts receiving no treatment served as control group. The photoaged cell model was verified by β-galactosidase staining, Western blot analysis for determining P21 protein expression, and cell counting kit-8 (CCK8) assay for evaluating cell viability. Moreover, Western blot analysis was performed to determine the protein expression of autophagy-related proteins P62, microtubule-associated protein 1 light chain 3 (LC3) -Ⅰand LC3-Ⅱ in photoaged human dermal fibroblasts, and real-time quantitative RCR (qRT-PCR) to verify the differential expression of circIGF2BP3 between photoaged and normal human dermal fibroblasts. Furthermore, circIGF2BP3 was biologically annotated. Some cultured primary human dermal fibroblasts were divided into 4 groups: empty vector group transfected with an empty vector, UVA + empty vector group transfected with an empty vector followed by repeated UVA radiation, circIGF2BP3 group transfected with a circIGF2BP3-overexpressing lentiviral vector, UVA + circIGF2BP3 group transfected with a circIGF2BP3-overexpressing lentiviral vector followed by repeated UVA radiation. Western blot analysis was performed to determine the expression of autophagy-related proteins. Statistical analysis was carried out by using t test, one-way analysis of variance and least significant difference- t test. Results:Compared with the control group, the UVA radiation group showed significantly increased proportions of β-galactosidase-positive cells (61.33% ± 5.78% vs. 6.37% ± 0.32%, t = 9.49, P < 0.01) and P21 expression (1.25 ± 0.03 vs. 1.00 ± 0.05, t = 4.26, P < 0.05), but significantly decreased cell viability (74.33% ± 3.48% vs. 100%, t = 7.38, P < 0.01). Moreover, the P62 expression and LC3-Ⅱ/Ⅰ ratio were significantly higher in the UVA radiation group than in the control group (both P < 0.05). The relative expression of circIGF2BP3 was 0.72 ± 0.04 in the photoaged human dermal fibroblasts, which was significantly lower than that in the normal human dermal fibroblasts (1.00 ± 0.03, t = 5.46, P < 0.01). The P62 expression and LC3-Ⅱ/Ⅰ ratio were significantly lower in the circIGF2BP3 group (0.60 ± 0.01, 0.71 ± 0.01, respectively) than in the empty vector group (1.00 ± 0.02, 1.00 ± 0.01, t = 16.25, 2.75, P < 0.01, < 0.05, respectively), and lower in the UVA + circIGF2BP3 group (1.05 ± 0.02, 2.04 ± 0.05, respectively) than in the UVA + empty vector group (1.31 ± 0.02, 2.72 ± 0.14, t = 10.493, 6.472, respectively, both P < 0.01) . Conclusion:circIGF2BP3 can regulate autophagy in UVA-induced photoaged dermal fibroblasts, which provides a new potential therapeutic target for the prevention and treatment of skin photoaging.

Objective: To investigate the degree and influencing factors of glycosylated hemoglobin ( HbA1c ) control among patients with type 2 diabetes ( T2DM ) living in rural communities of Zhejiang Province, so as to provide the reference for optimizing the management of diabetes patients in rural communities. Methods: Permanent adult patients with T2DM that were registered and received standardized management in all communities of Jiashan County and Suichang County, and 6 communities of Yongkang City, Zhejiang Province in 2016 were recruited, and their demographic characteristics were captured from the health record system and chronic disease management system. The height, body weight, waist circumstance and blood pressure were measured, and HbA1c and blood lipid parameters were detected. The degree of HbA1c control ( <7% ) was analyzed, and its influencing factors were identified using a multivariable logistic regression model. Results: A total of 10 339 patients with T2DM were enrolled, including 4 520 men ( 43.72% ) and 5 819 women ( 56.28% ), with a mean age of ( 63.54±9.78 ) years and the mean course of diabetes of ( 6.36±4.73 ) years. The rate of HbA1c control was 47.89%. Multivariable logistic regression analysis showed that males ( OR=1.123, 95%CI: 1.024-1.233 ), region ( Suichang, OR=2.413, 95%CI: 2.106-2.765; Yongkang, OR=2.460, 95%CI: 2.188-2.767 ), course of disease ( 5-9 years, OR=1.724, 95%CI: 1.504-1.977; 10 years and longer, OR=2.881, 95%CI: 2.477-3.351 ), use of hypoglycemic drugs ( OR=1.203, 95%CI: 1.089-1.329 ), development of chronic complications ( OR=1.190, 95%CI: 1.027-1.379 ), uncontrolled blood pressure ( OR=1.140, 95%CI: 1.030-1.261 ), uncontrolled blood lipid ( OR=1.258, 95%CI: 1.104-1.433 ), and smoking ( OR=1.318, 95%CI: 1.165-1.491 ) were statistically associated with HbA1c control among T2DM patients. Conclusion The rate of HbA1c control was 47.89% among T2DM patients in rural communities of Zhejiang Province. HbA1c control should be given a high priority among men living in low-economic-level regions with long course of disease, use of hypoglycemic drugs, chronic complications, smoking, uncontrolled blood pressure and lipid.