Objective:To identify genes interacting with the ubiquitin-conjugating enzyme E2S (UBE2S) in melanoma, and to explore the molecular mechanisms regulating the biological behavior of melanoma cells by UBE2S.Methods:Cultured A375 melanoma cells were divided into 2 groups to be transfected with LV-UBE2S-RNAi (14011-1) -containing lentivirus (gene knockdown group) and a negative control lentivirus CON053 (negative control group), respectively, and the UBE2S gene-knockdown cell line and negative control cell line were established. RNA was extracted from the two cell lines, purified, fragmented, and hybridized with GeneChip probes, followed by washing and dyeing, and then microarray data were obtained. Ingenuity pathway analysis (IPA) software was used to further analyze the gene expression profiling data to identify potential genes interacting with UBE2S. Western blot analysis was performed to verify the downstream molecules regulated by UBE2S. A two-tailed t test was used to screen for differentially expressed genes. Results:A total of 512 differentially expressed genes with a|fold change| > 2 and a P value < 0.05 were screened out between the gene knockdown group and negative control group, including 247 upregulated genes and 265 downregulated genes. IPA of differentially expressed genes revealed that interferon signaling and liver X receptor (LXR) /retinoid X receptor (RXR) activation pathways were significantly activated, while eukaryotic initiation factor-2 (EIF2) signaling and nuclear factor-κB (NF-κB) signaling pathways were inhibited. Among the upstream regulators, IFNA2 was predicted to be strongly activated, and NF-κB (complex) to be strongly inhibited. Based on the above bioinformatics analysis results, it was suggested that the UBE2S gene may exert some effects in melanoma cells by regulating the expression of IFITM1, STAT1, ISG15 and TNFRSF11B genes. Western blot analysis showed that IFITM1 protein was not expressed in A375 cells before and after the UBE2S gene knockdown. After the UBE2S gene knockdown, ISG15 protein expression was upregulated by 19.94 times, STAT1 protein expression was upregulated by 1.47 times, and TNFRSF11B protein expression was downregulated by 79.1%. Conclusion:UBE2S may interact with STAT1, ISG15 and TNFRSF11B to regulate the biological behavior of melanoma cells.

Objective:To evaluate the correlation between the timed up and go(TUG)test and fall risks in elderly frail patients.Methods:From July to September 2019, elderly frail patients who were treated at the cardiovascular department of our hospital were enrolled.Basic clinical data and fall-related information of patients were collected.Patients were divided into the fall group and the non-fall group.Results on the body mass index(BMI), TUG, 4-meter maximum walking speed(4 m MWS)and Barthel index were compared between the two groups.The correlation between TUG and each indicator was examined.Multivariate Logistic regression analysis was used to analyze the correlation between the TUG and falls in elderly patients.Results:A total of 96 eligible patients were enrolled, including 35 in the fall group and 61 in the non-fall group.The average TUG time was longer in the fall group than in the non-fall group(16.45±6.44 s vs.10.17±2.91 s, t=-6.556, P<0.001). The correlation analysis results showed that the TUG was correlated with falls and 4 m MWS( r=0.582 and 0.875, both P<0.001). Multivariate Logistic regression analysis showed that the TUG( OR=1.201, 95% CI: 1.111-1.470, P=0.004)and 4 m MWS( OR=1.146, 95% CI: 1.063-1.244, P=0.015)were risk factors for falls. Conclusions:The TUG is correlated with fall risks in elderly frail patients and should be recommended as a routine test in clinical practice.

Objective To investigate the positive rate of urine JC polyomavirus(JCV)and urine BK polyomavirus(BKV)in HIV patients,and to evaluate the relationship between urine JCV positive and renal function and lymphocyte count during long-term follow-up.Methods A total of 180 HIV-1 positive patients were enrolled in the Second Department of Infection,Xixi Hospital of Hangzhou,urine samples were collected for JCV and BKV detection.Renal function and CD4+cell count were collected at baseline before taking an-tiretroviral therapy(ART)and at 1,2,3,4,and 5 years of follow-up.Results The detection rates of JCV and BKV in urine of 180 HIV-1 positive patients were 25.55％(46/180)and 1.67％(3/180),respectively.Before ART,the total number of lymphocytes in urine JCV positive group was 1699.62±790.44/μl,which was lower than that in urine JCV negative group(2113.59±832.73/μl),and the difference was statistically significant(P=0.022).During ART initiation follow-up,serum creatinine levels in urine JCV posi-tive group were lower than those in urine JCV negative group.At the second year of follow-up,the serum creatinine level in urine JCV positive group was 70.00(64.00,77.00)μmol/L,which was lower than that in urine JCV negative group[76.00(67.00,85.00)μmol/L],and the difference was statistically significant(P=0.023).At the second year of follow-up,the glomerular filtration rate(GFR)of the urine JCV positive group was 118.00(106.25,133.75)ml/min,which was higher than that of urine JCV negative group[113.00(97.00,128.00)ml/min],and the difference was statistically significant(P=0.045).At the fourth year of follow-up,ser-um creatinine level in urine JCV positive group was 70.34±14.98μmol/L,which was lower than that in urine JCV negative group(76.6±14.45 μmol/L),and the difference was statistically significant(P=0.023).Conclusion The detection rates of JCV and BKV in urine of HIV-1 positive patients are negatively correlated with total lymphocyte levels.Urine JCV positive has a protective effect on the kidney,and urine JCV negative can be used as an early indicator of renal damage in HIV patients.

Purpose To summarize the clinical features, MRI features and pathological features of aggressive angiomyxoma (AAM) occurring in pelvic and perineal areas. Materials and Methods The clinical, MR and pathological data of 7 patients with AAM pathologically confirmed were retrospectively analyzed. Results Seven cases of AAM were all females, aged from 12 to 45 years old. Among them, 5 cases were treated due to perineal area tumors, and 2 cases due to relapses after surgical treatment of perineal area AAM in other hospitals. MRI findings: 7 cases were single occurrence with the maximum diameter of 5.9-15.7 cm; 6 cases were in irregular shape, with no envelope and unclear borders with adjacent tissues or organs; 1 case was in long column shape with clear boundary and pseudocapsule visible. On T1WI, AAM showed equal signal (4/7) or equal, low mixed signal (3/7). On T2WI, AAM showed high and low mixed signals with 5 cases of "vortex sign". 5 cases underwent routine enhancement examinations, all demonstrating significant enhancement, 4 cases of which displayed heterogeneous enhancement ("vortex sign" was detected in 2 cases) and 1 case homogeneous enhancement. On diffusion weighted imaging, AAM manifested as homogeneous (3/7) or heterogeneous (4/7) high signal. Pathological examination showed that a large amount of mucin-like matrix was contained in AAM, and there were scattered fusiform and astral tumor cells and abundant arterial and venous blood vessels seen locally. Tumor cells infiltrated the peripheral fat or muscle tissues at the border. Conclusion The characteristic MRI of AAM is manifested as "vortex sign", which corresponds to its pathological features such as sparse tumor cells, mucin-like matrix in large amount, and locally rich blood vessels, suggesting great significance for imaging diagnosis.

Objective To study the relationship between miR-655 with the clinical features of glioma and its role in diagnosis and prognosis.Methods The tissues and para cancerous tissues of 97 patients with glioma were collected from January 2013 to May 2017 at the Affiliated Hospital of Inner Mongolia Medical Universi-ty.The expression of miR 655 was detected by real-time fluorescence quantitative PCR,and the diagnosis and prognosis of miR-655 in glioma were analyzed.Results The expression of miR-655 in the cancerous tissues was significantly lower than that of the paracancerous tissues of the glioma (P<0.001),which was signifi-cantly related to the size of the tumor (P=0.024) and the WHO grading (P=0.002).COX regression analy-sis showed that the expression of miR-655 (P=0.004)and the degree of tumor resection(P=0.008) were in-dependent risk factors for glioma;ROC analysis showed that the area under the curve of the glioma (AUC) under the miR-655 expression was 0.959 (95% CI:0.936 -0.982);when the expression of miR-655 was 0.755,the sensitivity and specificity of the diagnosis were 89.7% and 88.7% respectively.The Kaplan Meier analysis of the expression of glioma was statistically significant (P=0.004).The lower expression of miR-655 indicated the worse the prognosis of glioma patients,the COX analysis miR-655 was an independent risk factor for the survival of glioma patients.Conclusion miR-655 can be used as a marker of poor prognosis in diagnosis and prognosis.

Objective:To observe the clinical efficacy of Qigui Tangtongning Granules in the treatment of diabetic peripheral neuropathy (DPN) with qi deficiency and blood stasis.Methods:Prospective cohort study. A total of 80 DPN patients with Qi deficiency and blood stasis in Endocrinology Department of the First Affiliated Hospital of Anhui University of Chinese Medicine from May 2021 to May 2022 who met the inclusion criteria were divided into 2 groups by random number table method, with 40 cases in each group. The control group was treated with epalrestat on the basis of routine hypoglycemia, and the treatment group was treated with Qigui Tangtongning Granules on the basis of control group. Both groups were treated for 8 weeks. TCM syndromes were scored before and after treatment. Disease severity was assessed using the Toronto Clinical Scoring System (TCSS). The motor nerve conduction velocity (MNCV) and sensory nerve conduction velocity (SNCV) of median nerve and common peroneal nerve were detected by electromyography/induced potentiometer. Serum CRP, TNF-α and IL-6 were detected by ELISA, fasting blood glucose (FPG) and two hours post-meal blood glucose (2 hPG) were detected by automatic biochemical analyzer, and glycosylated hemoglobin (HbA1c) was detected by automatic HBA1C analyzer. Adverse reactions were recorded and clinical efficacy was evaluated.Results:The total effective rate was 95.0% (38/40) in the treatment group and 77.5% (31/40) in the control group, the difference between the two groups was statistically significant ( χ2=5.17, P=0.023). After treatment, the TCM syndrome score and TCSS score of the treatment group were significantly lower than those in the control group ( t=-3.19 and -7.63, P<0.01); Median nerve SNCV [(47.90±4.51) m/s vs. (44.76±3.72) m/s, t=3.40], MNCV [(53.79±3.65) m/s vs. (51.32±4.25) m/s, t=2.79] and common peroneal nerve SNCV [(44.21±2.08) m/s vs. (40.51±2.49) m/s, t=7.23], MNCV [(44.63±4.72) m/s vs. (41.36±4.87) m/s, t=3.05] were significantly higher than those in the control group ( P<0.01); FPG [(5.05±0.63) mmol/L vs. (7.05±1.23) mmol/L, t=-9.17], 2 hPG [(9.10±1.64) mmol/L vs. (12.19±2.61) mmol/L, t=-6.35], HbA1c [(6.79±0.90) % vs. (7.22±1.02) %, t=-2.02] were significantly lower than those in the control group ( P<0.01 or P<0.05); TNF-α [(15.75±5.44) ng/L vs. (32.01±5.33) ng/L, t=-13.51], hs-CRP [(2.58±0.80) mg/L vs. (3.79±1.04) mg/L, t=-5.83], IL-6 [(18.20±4.92) ng/L vs. (29.97±5.18) ng/L, t=-10.41] were significantly lower than those in the control group ( P<0.01). No obvious adverse reactions were observed in 2 groups during treatment. Conclusion:Qigui Tangtongning Granules combined with conventional Western medicine can improve nerve conduction velocity, reduce inflammation and improve clinical efficacy in DPN patients with Qi-deficiency and blood-stasis syndrome.

Objective ToevaluatethevalueofMRDKIandDWIindiagnosingendometrialcarcinomaandevaluatingitspathologicalgrade. Methods TheDKIandDWIdataof48patientswithendometrialcarcinomaand27patientswithnormalendometrium wereanalyzed retrospectively.Thevaluesofmeankurtosis (MK),meandiffusion (MD)andADCinendometrialcarcinomaandnormalendometrium were measuredrespectively.Thesimilaritiesanddifferencesoftheparametersbetweentheendometrialnormalgroup(G0)andtheendometrialcarcinoma group (G1,G2,G3)werecomparedandanalyzed.TheROCcurvewasemployedtoevaluatethediagnosticefficacyandthresholdof eachparameter.P earson correlation wasappliedtoanalyzethecorrelationbetweeneachparametervalueandpathologicalgrade.Results The MK valuesincreasedgradually,meanwhiletheMDandADCvaluesdecreasedgraduallyinG0,G1,G2andG3groups.ExceptforMDand ADCvaluesbetweenG0andG1groups,othervalueswerestatisticallysignificantdifferent(P<0.05)betweendifferentgroups.In differentiatingoftheG0/(G1+G2+G3),G0/G1,G1/G2,G2/G3,theMKvalueshadthehighestdiagnosticefficacy(AUC=09.2,07.6,09.0,0.96, P<0 .05 ).M oreover ,in the correlation of pathological grading ,M K>M D>A D C (r=0 .850 ,0 .781 ,0 .709 ,P<0 .05 ).Conclusion Both DKIandDWIcandiagnoseandevaluatepathologicalgradeofendometrialcarcinoma.ComparedwithDWI,DKIembracesmoreperfectmathematical modelandmoresensitiveparameters,andcanbeusedasaneffectivemethodtoevaluatethepathophysiologicalfeaturesofendometrialcarcinoma.

Objective:To analyze the use and satisfaction of smart healthcare in public hospitals in Hangzhou from the perspective of patient experience.Methods:From August to September 2020, 501 patients from 2 class A tertiary hospitals and 5 community health centers in Hangzhou were randomly selected for questionnaire survey. The survey included their usage and satisfaction of smart healthcare. Based on SERVQUAL theory, the patient satisfaction scale included 5 dimensions of tangibility, reliability, reactivity, assurance, and empathy. The factors affecting patient satisfaction were analyzed by single factor and multi factor analysis.Results:The average score of patient satisfaction was 3.94±0.56. The satisfaction with tangibility(4.25±0.64), assurance(4.07±0.71)and reactivity(4.02±0.73)was higher, while the scoring of empathy(3.84±0.77)and reliability(3.52±0.70)were lower than the average score. The results of multiple linear regression analysis showed that patients′ educational level, occupational type and UTAUT score had a certain impact on satisfaction( P<0.05). Conclusions:The overall satisfaction of patients with smart healthcare in Hangzhou is high. In the future, we should pay more attention to big data security and personal privacy protection, upgrade the smart medical system, improve the hospital Internet platform, empower smart healthcare service system through digital technology, and promote the comprehensive digital transformation of health.

Objective To determine the expression of ubiquitin-conjugating enzyme E2S (UBE2S) in malignant melanoma (MM),and to evaluate its effect on the biological behavior of melanoma cells.Methods Immunohistochemical study was performed to determine the UBE2S expression in 128 primary MM tissue chips,64 metastatic MM tissue chips,16 non-tumor tissue chips (8 paralesional normal skin tissues and 8 normal epidermal tissues).Real-time quantitative RCR was conducted to determine the UBE2S mRNA expression in the melanoma cell lines A375,MUM-2B and MUM-2C.The melanoma cell lines A375 and MUM-2B were divided into 2 groups separately:interference group transfected with a lentiviral vector carrying UBE2S RNA interference sequence,and control group transfected with a lentiviral vector carrying control sequence.After 72 hours,real-time quantitative RCR was performed to determine the UBE2S mRNA expression in the melanoma cell lines A375 and MUM-2B.Caspase-3/7 activity in the groups was assessed by using kits,and cell apoptosis and cell cycle distribution were detected by flow cytometry.The effect of UBE2S knockdown on the migratory and invasive abilities of and N-cadherin expression in A375 cells were evaluated by Transwell assay and Western blot analysis respectively.Statistical analysis was carried out with SPSS 22.0 software by using independent sample t-test for the comparison of normally distributed data between two groups,chi-square test for enumeration data,MannWhitney U test for the comparison of non-normally distributed data,and Spearman's coefficient for assessment of the correlation of UBE2S expression with T staging of melanoma.Results UBE2S was highly expressed in 98 (51.0％) MM tissues,but lowly expressed in 16 non-tumor tissues,and the UBE2Sexpression rate significantly differed between the above two kinds of tissues (x2 =11.905,P ＜ 0.01).UBE2S expression was negatively correlated with T staging of melanoma (ρ =-0.210,P =0.043).The relative mRNA expression of UBE2S significantly differed among the A375,MUM-2B,and MUM-2C cells (F =817.228,P ＜ 0.01).After UBE2S knockdown,the caspase-3/7 activity was significantly up-regulated in the A375 interference group (t =17.572,P ＜ 0.01) and MUM-2B interference group (t =24.552,P ＜0.01) compared with the A375 and MUM-2B control groups respectively.Compared with the control group,the A375 interference group showed significantly increased proportion of A375 cells at G1 phase (t =7.365,P ＜ 0.01),decreased proportion at S phase (t =-9.190,P ＜ 0.01),and no change in the proportion of A375 cells at G2/M phase (t =-0.227,P ＞ 0.05).The MUM-2B interference group showed significantly increased proportions of MUM-2B cells at G1 (t =12.676,P ＜ 0.01) and G2/M phases (t =13.045,P ＜0.01),but significantly decreased proportion at S phase (t =-15.718,P ＜ 0.01) compared with the control group.Transwell assay revealed decreased migratory and invasive abilities of A375 cells in the interference group compared with the control group (t =-35.727,-125.000,P ＜ 0.05,＜ 0.01,respectively).Western blot analysis showed down-regulated expression of N-cadherin protein in A375 cells in the interference group compared with the control group.Conclusions UBE2S is over-expressed in melanoma tissues,whose expression is associated with the T staging of melanoma.Knockdown of UBE2S affects the apoptosis,cell cycle,migration and invasion of melanoma cells,and may promote the metastasis of MM cells by regulating N-cadherin expression.

Objective We investigated the effects of Xueguan Ruanhua Pills(XGRHW) on ferroptosis in ApoE-/- atherosclerotic mice through the nuclear factor E2 related factor 2 (Nrf2)/xCT/glutathione peroxidase 4 (GPX4) signaling pathway.Methods Ten male C57BL/6J mice in the normal group were fed normal chow. Additionally, 50 ApoE-/- mice were fed high-fat chow for 12 weeks, and were divided into the following five groups (10 mice per group): the model group, the XGRHW low-dose (2.34g/kg) group, the XGRHW high-dose (4.68 g/kg) group, the XGRHW high-dose combined with the Nrf2 inhibitor ML385 (0.03 g/kg) group, and the ferrostatin-1 (1 mg/kg) group. Drugs were administered for 6 weeks. The blood levels of four types of lipids were detected by an automatic lipid analyzer, lipid deposition in the aorta was observed by Oil Red O staining, histomorphological changes in the aortic sinus were observed by HE staining, the serum levels of Fe2+, MDA, GSH, and SOD were determined by colorimetric assays, and the expression levels of FTH1 and FTL in the aortic sinus were observed by immunofluorescence. The protein levels of Nrf2, xCT, and GPX4 in mouse aortic tissues were detected by Western blotting. The ultrastructural changes of aortic mitochondria were observed by transmission electron microscopy.Results Compared with the normal group, mice in the model group showed obvious lipid plaque deposition in the aorta, severely calcified lesions in the aortic sinus, elevated serum levels of TC, TG, LDL-C, Fe2+, and MDA, decreased levels of HDL-C, SOD, and GSH (P<0.01), and decreased protein expressions of aortic Nrf2, xCT, and GPX4 as well as the iron storage proteins FTH1 and FTL (P<0.01), and serve damage to mitochondrial structure and morphology. Compared with the model group, the relative aortic plaque area was decreased, calcified lesions in the aortic sinus were decreased, serum levels of TC, TG, LDL-C, Fe2+, and MDA were decreased, and HDL-C, SOD, and GSH levels were increased in the XGRHW low-dose and high-dose and ferrostatin-1 groups (P<0.05 or P<0.01), and Nrf2, xCT, GPX4, and the iron storage proteins FTH1 and FTL were upregulated in aortic tissues (P<0.05 or P<0.01), and mitochondrial structure approaching normal. In the XGRHW high-dose+ML385 group, compared with the XGRHW high-dose group, the levels of blood lipids and lipid peroxidation were increased and the protein levels of Nrf2, xCT, and GPX4 in aortic tissue and the iron storage proteins FTH1 and FTL were decreased (P<0.01), and mitochondrial structure was damaged indicating that ML385 could inhibit the therapeutic effect of the XGRHW in atherosclerotic mice.Conclusion The XGRHW can improve blood lipid levels and reduce the degree of arterial plaque lesions in atherosclerotic mice, and its mechanism of action may be related to activation of the Nrf2/xCT/GPX4 pathway to inhibit ferroptosis.