[Long-term outcomes of lower extremity arteriovenous grafts].

Objective: To evaluate the long-term outcomes of lower extremity arteriovenous graft (AVG) in hemodialysis patients. Methods: Hemodialysis patients with lower extremity AVG from August 2015 to July 2023 in the Department of Vascular Surgery, Longhua Hospital, Shanghai University of Traditional Chinese Medicine were enrolled. Therapeutic effects and complications of AVG were retrospectively analyzed. Results: A total of 83 cases aged (58.9±13.3) years were enrolled, including 25 males and 58 females. The success rate of the operation was 100% (83/83), and no perioperative complications occurred. The follow-up time [M (Q1, Q3)] was 38.4 (22.6, 55.3) months, with a follow-up rate of 92.8% (77/83). There were 9 cases (11.7%) of puncture site infection, 5 cases (6.5%) of pseudoaneurysm, 2 cases (2.6%) of seroma, 3 cases (3.9%) of lower limb ischemia, 48 cases (62.3%) of stenosis and 24 cases (31.2%) of thrombosis during the follow-up period. The 6-month, 1-year, 2-year, 3-year and 5-year primary patency rates after surgery were 78.9%, 61.2%, 39.0%, 27.0% and 16.3%, respectively, assisted primary patency rates were 93.5%, 82.5%, 74.9%, 68.0% and 53.0%, respectively, and secondary patency rates were 96.1%, 94.7%, 93.1%, 91.3% and 75.3%, respectively. Conclusion: For patients whose vascular resources of upper limbs are exhausted, lower extremity AVG is a safe and effective hemodialysis vascular access.

Causal association between aspirin use and risk of endometrioid carcinoma: a Mendelian randomization study.

OBJECTIVE: The aim of the study was to investigate the causal relationship between aspirin use and the risk of endometrial endometrioid cancer (EEC) using two-sample Mendelian randomization (TSMR) and multivariable Mendelian randomization (MVMR) study. MATERIALS AND METHODS: A TSMR analysis was conducted to estimate the potential causal relationship between aspirin use and the risk of EEC using genome-wide data from Genome-wide association study (GWAS). The causal association between aspirin use and EEC was further analyzed by MVMR analysis after adjusting for various factors such as obesity, hypertension, diabetes, and infertility. The single nucleotide polymorphism (SNP) data associated with aspirin use and EEC was obtained from the GWAS catalog database. RESULTS: A total of six SNPs were included as instrumental variables in TSMR, which showed that taking aspirin reduced the risk of EEC [OR = 0.02, 95% CI = 0-0.28, p = 0.005, inverse variance weighted (IVW) method]. Besides, the results of the weighted median (WME) method, weighted mode, and simple mode were consistent with the results shown by the IVW method. After further using the MVMR method, the causal association of aspirin use and prevention of EEC onset remained significant after adjusting for the effects of obesity, hypertension, and diabetes (OR = 0.076, 95% CI = 0.007-0.793, p = 0.031). Sensitivity analyses, including heterogeneity, horizontal multiplicity, and leave-one-out tests, showed the reliability of the instrumental variables, proving that the results were reliable and not significantly biased. CONCLUSIONS: Taking aspirin can reduce the risk of EEC morbidity, and it is expected to be of great significance for the early prevention and treatment of endometrial cancer by exploring the biological mechanism of aspirin on endometrioid cancer at a deeper level.

The GluN2B-Containing NMDA Receptor Alleviates Neuronal Apoptosis in Neonatal Hypoxic-Ischemic Encephalopathy by Activating PI3K-Akt-CREB Signaling Pathwa.

Neonatal hypoxic-ischemic encephalopathy (HIE) is a disease caused by insufficient blood supply in the brain in newborns during the perinatal period. Severe HIE leads to patient death, and patients with mild HIE are at increased risk of cognitive deficits and behavioral abnormalities. The NMDA receptor is an important excitatory receptor in the central nervous system, and in adult hypoxic-ischemic injury both subtypes of the NMDA receptor play important but distinct roles. The GluN2A-containing NMDA receptor (GluN2A-NMDAR) could activate neuronal protective signaling pathway, while the GluN2B-NMDAR subtype is coupled to the apoptosis-inducing signaling pathway and leads to neuronal death. However, the expression level of GluN2B is higher in newborns than in adults, while the expression of GluN2A is lower. Therefore, it is not clear whether the roles of different NMDA receptor subtypes in HIE are consistent with those in adults. We investigated this issue in this study and found that in HIE, GluN2B plays a protective role by mediating the protective pathway through binding with PSD95, which is quite different to that in adults. The results of this study provided new theoretical support for the clinical treatment of neonatal hypoxic ischemia.

[Efficacy and prognostic analysis of orthopedic surgery in patients with newly diagnosed multiple myeloma].

Objective: To evaluate the efficacy and prognosis of orthopedic surgical resection surgery in patients with newly diagnosed multiple myeloma (NDMM). Methods: This retrospective cohort study collected clinical data of patients with NDMM who underwent surgery due to spinal cord compression or pathological long-bone fractures at the Peking Union Medical College Hospital from 1 January 2003 to 31 December 2021. Patients who received biopsy or vertebroplasty/kyphoplasty were excluded and patients with the same degree of bone disease and who did not undergo any surgical intervention were selected as controls. Visual analogue scale (VAS) and physical status (ECOG) scores, progression-free survival (PFS), and overall survival (OS) were compared. Statistical analysis included the χ2-test, t-test, and Kaplan-Meier methods. Results: Baseline data were compared between the surgical group (n=40 with 43 interventions) and the non-surgical group (n=80), and included sex, age, paraprotein type, International Staging System (ISS), number of lytic lesions, cytogenetic abnormalities, first-line treatment, and the proportion of patients receiving autologous stem cell transplantation (ASCT) (all P>0.05). Serum M protein levels in the surgical group were significantly lower than those of the non-surgical group [(21.95±16.44) g/L vs. (36.18±20.85) g/L, P=0.005]. The surgical lesions involved the axial skeleton (79.1%, 34/43) or the extremities (20.9%, 9/43). VAS and ECOG scores improved significantly after surgery (VAS: 2.30±0.80 vs. 6.60±1.50, P<0.001; ECOG: 2.09±0.59 vs. 3.09±0.73, P<0.001). The median follow-up time was 51 months. Kaplan-Meier survival analysis suggested that the median PFS (25 vs. 29 months) and OS (46 vs. 60 months) were comparable between the surgical and non-surgical intervention groups (both P>0.05). Subgroup analysis showed that among patients with ISS Ⅰ or those who had received ASCT, PFS in the surgical group was similar to that of the non-surgical intervention group (both P>0.05), while OS was worse (P=0.005, 0.017). Patients with ISS Ⅱ/Ⅲ scores or without ASCT had similar PFS and OS between the surgical and non-surgical intervention groups (all P>0.05). Cox multivariate analysis suggested that ISS and ASCT were independent prognostic factors for OS (ISS: HR=0.42, 95%CI 0.19-0.93, P=0.031; ASCT: HR=0.41, 95%CI 0.18-0.97, P=0.041), while orthopedic surgery did not influence survival (P=0.233). Conclusion: For patients with NDMM, orthopedic surgical resection decreased bone-related complications and improved quality of life, but did not affect survival.

[Optimal surveillance intensity of cystoscopy in intermediate-risk non-muscle invasive bladder cancer].

OBJECTIVE: To determine the optimal cystoscopic frequency for intermediate-risk non-muscle invasive bladder cancer. METHODS: Patients with intermediate-risk non-muscle invasive bladder cancer, who underwent transurethral resection of bladder tumor in Peking University People's Hospital from January 2001 to October 2019, were retrospectively analyzed. Their clinical, pathological and follow-up data were collected. In postoperative 2-year period, the patients were underwent cystoscopy every 3 to 6 months. Depending on recurrence and progression of the patients, we hypothesized three strategies of surveillance intensity in the first 2 years after surgery: model 1: 3-month intervals, model 2: 6-month intervals, and model 3: 12-month intervals. The differences in the numbers and time of delayed detection of recurrence and progression were compared among the three models. RESULTS: A total of 185 patients were enrolled, including 144 males (77.8%) and 41 females (22.2%). The median age was 68 (59-76) years. There were 118 cases (63.8%) with single tumor and 67 cases (36.2%) with multiple tumor. Of the patients 179 (96.8%) had stage Ta and 6 (3.2%) had stage T1. There were 108 cases (58.4%) with high-grade disease and 77 cases (41.6%) with low-grade disease. During the follow-up period of the first 2 years, 52 patients (28.1%) had recurrence, 133 cases (71.9%) had no recurrence, 11 cases (5.9%) had progression and 174 cases (94.1%) had no progression. Compared with model 1, 29 (55.8%) delayed detection of recurrence in model 2 vs. 41 (78.8%) delayed detection of recurrence in model 3, and the difference was statistically significant (P=0.012). The median delayed time of detecting recurrence was 1.00 months in model 1, 1.99 months in model 2 and 4.19 months in model 3, respectively. There were statistically significant differences between mode 1 and model 3 (P=0.001), and between model 2 and model 3 (P=0.013). Compared with model 1, 5 (45.4%) delayed detection of progression in model 2 vs. 8 (72.7%) delayed detection of progression in model 3, and the difference was not statistically significant. The median delayed time of detecting progression was 1.00 month in model 1, 2.00 months in model 2 and 3.00 months in model 3, respectively. There was no statistically significant difference among them. CONCLUSION: Although providing slightly slower detection of tumor recurrence and progression, compared with 3-month intervals of cystoscopy, 6-month intervals do not result in serious adverse outcomes and reduce cost and pain of the patients, which is feasible in intermediate-risk non-muscle invasive bladder cancer.

[Establishment of a mutation prediction model for evaluating the efficacy of immunotherapy in renal carcinoma].

OBJECTIVE: To establish a mutation prediction model for efficacy assessment, the genomic sequencing data of renal cancer patients from the MSKCC (Memorial Sloan Kettering Cancer Center) pan-cancer immunotherapy cohort was used. METHODS: The genomic sequencing data of 121 clear cell renal cell carcinoma patients treated with immune checkpoint inhibitors (ICI) in the MSKCC pan-cancer immunotherapy cohort were obtained from cBioPortal database (http://www.cbioportal.org/) and they were analyzed by univariate and multivariate Cox regression analysis to identify mutated genes associated with ICI treatment efficacy, and we constructed a comprehensive prediction model for drug efficacy of ICI based on mutated genes using nomogram. Survival analysis and time-dependent receiver operator characteristic curves were performed to assess the prognostic value of the model. Transcriptome and genomic sequencing data of 538 renal cell carcinoma patients were obtained from the TCGA database (https://portal.gdc.cancer.gov/). Gene set enrichment analysis was used to identify the potential functions of the mutated genes enrolled in the nomogram. RESULTS: We used multivariate Cox regression analysis and identified mutations in PBRM1 and ARID1A were associated with treatment outcomes in the patients with renal cancer in the MSKCC pan-cancer immunotherapy cohort. Based on this, we established an efficacy prediction model including age, gender, treatment type, tumor mutational burden (TMB), PBRM1 and ARID1A mutation status (HR=4.33, 95%CI: 1.42-13.23, P=0.01, 1-year survival AUC=0.700, 2-year survival AUC=0.825, 3-year survival AUC=0.776). The validation (HR=2.72, 95%CI: 1.12-6.64, P=0.027, 1-year survival AUC=0.694, 2-year survival AUC=0.709, 3-year survival AUC=0.609) and combination (HR=2.20, 95%CI: 1.14-4.26, P=0.019, 1-year survival AUC=0.613, 2-year survival AUC=0.687, 3-year survival AUC=0.526) sets confirmed these results. Gene set enrichment analysis indicated that PBRM1 was involved in positive regulation of epithelial cell differentiation, regulation of the T cell differentiation and regulation of humoral immune response. In addition, ARID1A was involved in regulation of the T cell activation, positive regulation of T cell mediated cyto-toxicity and positive regulation of immune effector process. CONCLUSION: PBRM1 and ARID1A mutations can be used as potential biomarkers for the evaluation of renal cancer immunotherapy efficacy. The efficacy prediction model established based on the mutation status of the above two genes can be used to screen renal cancer patients who are more suitable for ICI immunotherapy.

[Bipolar androgen therapy followed by immune checkpoint inhibitors in metastatic castration resistant prostate cancer: A report of 4 cases].

The relationship between androgen and prostate cancer treatment has plagued the field of urologic oncology. To investigate the efficacy and safety of bipolar androgen therapy (BAT) followed by immune checkpoint inhibitor therapy in patients with metastatic castration resistant prostate cancer (mCRPC). In August 2020, Beijing Hospital conducted an investigator-initiated study: Bipolar androgen therapy followed by immune checkpoint inhibitor therapy in metastatic castration resistant prostate cancer. Up to now, the study has included 4 patients who completed the entire cycle of treatment. The mean age of the patients was 74.5 (68 to 82) years old, the mean prostate-specific antigen (PSA) was 20.8 (9.9 to 8.36) µg/L, the mean testosterone was 0.50 (0.00 to 1.81) µg/L, and the Gleason score were 10 and 9, 7, 7 respectively. The pain scale score before treatment was 1.5 (1 to 2). In this study, 4 patients completed the entire cycle of treatment, and the treatment effect of the patients showed great heterogeneity. PSA in case 1 decreased from 24.0 µg/L to 0.47 µg/L, testosterone increased from 0.175 6 µg/L to 2.62 µg/L. PSA in case 2 increased from 9.939 µg/L to 168.536 µg/L, and testosterone increased from 0.0 µg/L increased to 2.85 µg/L. PSA increased from 13.31 µg/L to 39.278 µg/L in case 3, testosterone increased from 0.0 µg/L to 2.54 µg/L. and PSA increased from 36.0 µg/L to 350.2 µg/L in the case 4, testosterone increased from 1.81 µg/L to 3.85 µg/L. Except for one patient who showed significant PSA remission, the PSA levels of the remaining three patients remained high overall. There were no adverse reactions reported in 4 patients. In the follow-up, case 1 continued to use PD-1 monoclonal antibody (median progression free survival time was 10 months). Two patients who had previously been resistant to enzalutamide received enzalutamide again after the whole cycle of treatment, and their PSA decreased again, which indicated that the patient was sensitive to enzalutamide again. BAT had a certain therapeutic effect on mCRPC patients, and the safety was controllable. Its tumor control effect still needed long-term follow-up verification in large-sample clinical trials. BAT has a certain therapeutic effect on mCRPC patient, especially the resensitivity of tumors to enzalutamide can be restored. Immune checkpoint inhibitors may have therapeutic potential in patients with prostate cancer treated with BAT and enzalutamide.

[Changes of serum lipid profiles and the correlation analysis in clinic-pathological features of pancreatic cancer patients].

Objective: To examine the characteristics of blood lipid profile and the correlation with clinic-pathological features of pancreatic cancer patients. Methods: The clinical and pathological data of 265 pancreatic cancer patients who received radical surgical treatment at Department of General Surgery,Qilu Hospital,Shandong University from January 2013 to September 2020 were collected and analyzed retrospectively. Among the 265 pancreatic cancer patients,there were 170 males and 95 females,with age of (61.0±9.6)years(range:28 to 86 years). General information,lipid indicators and clinic-pathological information were collected from electronic medical record system,and follow-up information gained by telephone. According to level of serum lipid in pancreatic cancer patients,265 patients were divided into dyslipidemia group(n=115) and normal lipid group(n=150). Pearson χ2,Student's t tests, variance analysis or univariate Logistic regression was used to analyze the correlation between dyslipidemia and clinico-pathological characteristics of pancreatic cancer,respectively. Kaplan-Meier survival curve was used to assessed the influence of dyslipidemia on prognosis of pancreatic cancer patients. Results: In 265 pancreatic cancer patients,115(43.4%)of them had dyslipidemias,and the most common form was increase of triglyceride(TG)(72.2%). In pancreatic cancer with dyslipidemias group,patients with body mass index ≥25 kg/m2 had higher proportion than normal lipid group(36.1%(26/72) vs. 21.2%(21/99),χ²=4.643,P=0.031); The proportion of carcinoma located at head of pancreas(83.5%(96/115) vs. 40.7%(61/150),χ²=49.412,P<0.01), staging of T1/T2(79.1%(91/115) vs. 60.7%(91/150),χ²=10.316,P<0.01) and lymphatic metastasis(36.5%(42/115) vs. 22.7%(34/150),χ²=6.007,P<0.01) were higher. In patients of pancreatic cancer, dyslipidemias were closely associated with tumor location(OR=10.529,P<0.01)and body mass index(OR=3.671,P=0.008). Serum lipid profile results showed that TG,total cholesterol and high-density lipoprotein(HDL) disorders were associated with tumor location(P<0.05). TG disorder had association with body mass index(P<0.05), and HDL disorder had association with tumor stage(P<0.05). Moreover, the result of survival analysis showed that dyslipidemia was not a factor to impact the prognosis of pancreatic cancer patients underwent surgery(P>0.05). Conclusions: In pancreatic cancer patients,TG disorder was the most common type of dyslipidemia. Dyslipidemia has closely association with clinicopathologic features,including tumor location,body mass index,tumor stage. However,dyslipidemia had little effect on prognosis of pancreatic cancer patients.

Whole-genome resequencing reveals genetic structure and introgression in Pudong White pigs.

Pudong White (PDW) pigs, historically originating from Shanghai, are the only Chinese indigenous pigs characterised by their completely white coats, with the exception of Rongchang pigs. However, there is limited information concerning their overall genetic structure or relationship with other breeds, especially the East Chinese (ECN) and European pigs. To uncover the genetic structure, selection signatures, and potential exotic introgression in PDW pigs, we sampled 15 PDW pigs using whole-genome sequencing (~20×). We then conducted in-depth population genetic analyses in 320 pigs from 27 global pig groups, namely, European wild boars, Chinese wild boars, and outgroup. Neighbour-joining tree and principal component analysis confirmed that PDW pigs belonged to the ecotype of ECN pigs. Both f3, D-statistics, and structure analysis showed that PDW pigs shared apparent alleles with Large White (LW) pigs. Three statistics, rIBD, a haplotype heat map and copy number variation, further indicated that PDW pigs shared apparent alleles with LW pigs at the KIT Proto-Oncogene, Receptor Tyrosine Kinase (KIT) and PARG-MARCHF8 loci, suggesting that the lineage of European pigs in PDW originated from LW pigs. After further detecting the KIT mutations in different pig breeds, PDW was confirmed to have the same duplication region 1, duplication region 2, and the splicing mutation on intron 17 of KIT as LW pigs that determine the white coat colour phenotype in European white pigs. We hypothesised that LW pigs were imported to China ∼110-160 years ago according to the admixture time estimate and then crossed with ECN pigs, resulting in the introgression of the KIT alleles that produce the white coat colour phenotype in the PDW pig breed. To our knowledge, this study presents the first thorough description of the genetic structure of PDW pigs via whole-genome resequencing data; moreover, the results provide a basis for the national project for the conservation of this unique Chinese local population.

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

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

[The association between blood homocysteine level and Chinese Healthy Eating Index in a suburb population of Shanghai].

Objective: To explore the relationship between the level of blood homocysteine (Hcy) and the total score of Chinese Healthy Eating Index (CHEI) and its item score. Methods: The subjects were recruited from the East China Natural Population Cohort Study, led by the School of Public Health in Fudan University, which was conducted in Zhongshan Community, Songjiang District of Shanghai from April to September 2017. By using the cluster random sampling method, 8 neighborhood committees were randomly selected from 18 neighborhood committees in Zhongshan community (Beimen, Baiyun, Dongwai, Huaqiao, Lantian village 1, Lantian village 2, Lantian village 4, and Lantian village 5). All the residents who met the standard and had lived in Shanghai for more than half a year were selected as research subjects. 4 995 subjects with complete survey information were finally included in this study. General information (age, sex, disease history, etc.), lifestyle (smoking, drinking, tea drinking, physical activity, etc.), food frequency and blood Hcy concentration were collected through questionnaire survey, physical examination and biological sample detection. The multivariate linear regression model was used to analyze the correlation between blood Hcy concentration and the total score of CHEI and its item score, and the multivariate logistics regression model was used to analyze the correlation between hyperhomocysteinemia (hHcy) and the total score of CHEI and its item score. Results: The age of the subjects was (56.72±9.72) years. The proportion of females, people with middle and high school education and high physical activity was 64.90% (3 241), 50.80% (2 539) and 63.20% (3 157), respectively. The blood Hcy concentration was (11.25±4.90) µmol/L, and the total prevalence of hHcy was 9.3% (467 cases). The results of multivariate linear regression showed that after adjusting for the relevant confounding factors, the blood Hcy concentration of subjects decreased with the increase of the total score of CHEI and the item score of fruit, milk, seafood, poultry and egg, but increased with the increase of the item score of total grain and tuber. In males, blood Hcy levels decreased with the increase of the item score of seafood and poultry [ß (95%CI) values were -0.343 (-0.582, -0.102) and -0.225 (-0.402, -0.046), respectively]. In females, the blood Hcy level decreased with the increase of the total score of CHEI and its item score of milk, egg, seafood and poultry [ß (95%CI) values were -0.130 (-0.207, -0.052), -0.091 (-0.148, -0.034), -0.016 (-0.026, -0.007), -0.069 (-0.122, -0.016), and -0.087 (-0.157, -0.017), respectively]. The results of multivariate logistic regression showed that the higher the total score of CHEI and its item score of milk and seafood, the lower the risk of hHcy [OR (95%CI) value were 0.986 (0.978, 0.995), 0.915 (0.864, 0.969), and 0.862 (0.806, 0.922), respectively]. In females, the higher the total score of CHEI and its item score of milk and seafood, the lower the risk of hHcy [OR (95%CI) values were 0.984 (0.970, 0.999), 0.877 (0.802, 0.958), and 0.845 (0.760, 0.941), respectively]. In males, the higher the total score of CHEI and its item score of seafood, the lower the risk of hHcy [OR (95%CI) values were 0.988 (0.977, 0.998) and 0.858 (0.791, 0.930), respectively]. Conclusion: The dietary pattern of residents in Zhongshan Community, Songjiang District, Shanghai can affect their own blood Hcy concentration and the risk of hHcy. The total score of CHEI and the item score of fruit, milk, seafood, poultry and eggs play an important role in reducing the level of blood Hcy. The higher the total score of CHEI and the item score of milk and seafood, the lower the risk of hHcy.

Asiatic Acid Induces Mitochondrial Apoptosis via Inhibition of JAK2/STAT3 Signalling Pathway in Human Osteosarcoma.

Osteosarcoma (OS), a severe malignant bone tumour, usually occurs in adolescents and children and has a poor prognosis. Asiatic acid (AA), an active component isolated from Centella asiatica (L.) Urb., exhibits appreciable anti-oxidant and anti-tumour activities. So far, the effects and underlying mechanisms of AA against OS have not been clarified. Here, we explored the anti-tumour effects of AA against human OS and the involved mechanism mediating its actions. To evaluate effects of AA on the cell proliferation of human OS cells, cell viability and colony formation assays were performed. Flow cytometry was used to evaluate apoptosis in OS cells exposed to AA and mitochondrial membrane potential. Western blotting and RT-PCR were applied to determine expression of the relevant proteins and their mRNA levels. Our explorations showed that AA inhibits proliferation of human OS cells in a concentration- and time-dependent manner, and induces apoptosis of OS cells by the intrinsic (mitochondrial) pathway. Importantly, we found that inhibition of the AA-induced phosphorylation of JAK2/STAT3 signalling molecules and the decrease in MCL-1 contributed to the anti-tumour efficacy of AA. Collectively, our results suggest that AA could evoke mitochondrial- induced apoptosis in human OS cells by suppression of the JAK2/STAT3 pathway and MCL-1 expression. These results strongly demonstrate that AA could be a potential anti-tumour agent for OS treatment.

MiR-16 inhibits proliferation of cervical cancer cells by regulating KRAS.

OBJECTIVE: The aim of this study was to explore the effects of micro ribonucleic acid (miR)-16 on the proliferation and apoptosis of cervical cancer (CC) cells and its related regulatory mechanism. MATERIALS AND METHODS: The downstream regulatory targets of miR-16 were analyzed based on the miRNA online database. HCC94 cells were selected as experimental objects. Subsequently, the cells were transfected with miR-16 mimic (miR-16 mimic group), miR-16 small interfering RNA (siRNA) (miR-16 siRNA group) and only Lipofectamine 2000 transfection reagent [blank control group and miR-16 normal control (NC) group]. The expression level of miR-16 in HCC94 cells was measured via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Cell counting kit-8 (CCK-8) assay, 5-Ethynyl-2'-deoxyuridine (EdU) staining assay and flow cytometry were then conducted to detect the effects of miR-16 on the viability, proliferation and apoptosis of HCC94 cells, respectively. Additionally, the effect of miR-16 on the protein expression level of Kirsten rat sarcoma viral oncogene homolog (KRAS) in HCC94 cells was determined via Western blotting. RESULTS: MiRNA online database analysis showed that KRAS was the downstream target of miR-16. Compared with miR-16 NC group, the viability and proliferation ability of HCC94 cells increased significantly in miR-16 siRNA group but decreased significantly in miR-16 mimic group (p<0.05). However, the apoptosis rate evidently declined in miR-16 siRNA group while increased remarkably in miR-16 mimic group (p<0.05). In addition, the protein expression level of KRAS in HCC94 cells was significantly higher in miR-16 siRNA group but significantly lower in miR-16 mimic group when compared with miR-16 NC group (p<0.05). CONCLUSIONS: MiR-16 is lowly expressed in HCC94 cells. Moreover, highly expressed miR-16 represses the viability and proliferation of HCC94 cells and promotes their apoptosis by targeted regulation on KRAS.

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

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

[Long-term analysis of safety and efficacy of standard percutaneous nephrolithotomy in patients with solitary kidneys].

OBJECTIVE: To evaluate the long-term efficacy and safety of ultrasound-guided percutaneous nephrolithotomy (PCNL) in the treatment of patients with solitary kidney stones. METHODS: The clinical data of 22 patients with solitary kidney stones treated with PCNL in Peking University People's Hospital from September 2008 to June 2014, with the follow-up data of more than 5 years were analyzed retrospectively. Perioperative indicators, postoperative stone free rate (SFR) and incidence of complications were recorded. Ultrasonography was used to evaluate the long-term stones recurrence rate. Serum creatinine and estimated glomerular filtration rate (eGFR) were used to assess the long-term renal function. RESULTS: In this group of 22 patients, the average age was (50.3±11.8) years, with 10 cases of anatomic solitary kidneys, 12 functional solitary kidneys, and the median stone diameter was 1.65 (1.1-3.9) cm. All the patients had multiple stones, including 7 cases of staghorn stones. The median pre-operative serum creatinine was 104.5 (60.0-460.0) µmol/L, and the mean eGFR was (60.3±29.4) mL/min, showing no statistically significant difference compared with that before surgery. The mean operative time was (88.2±42.0) min, and there were 11 cases of single-channel and double-channel PCNL. The median serum creatinine on the first day after surgery was 102.0 (63.0-364.0) µmol/L, and the mean eGFR was (58.0±25.1) mL/min. The mean postoperative hospital stay was (8.7±5.2) days. In this group, 5 patients (22.7%) presented short-term complications, among which 4 patients presented postoperative infection and massive hemorrhage at the same time, which improved after conservative treatment, and 1 patient presented pleural injury and improved after closed thoracic drainage. Two patients (9.1%) developed long-term complications, and ureteral stricture occurred 3 months after operation, which improved after balloon dilatation. The median follow-up time was 6.2 (4.7-11.1) years. The median serum creatinine at the last follow-up was 104.0 (72.4-377.0) µmol/L, and the mean eGFR was (60.1±23.7) mL/min, showing no statistically significant difference compared with that before surgery. Renal function decreased in 6 patients (27.3%). Initial and final SFR were 72.7% and 100%, respectively. During the 6.2-year follow-up, 9 patients (40.9%) experienced recurrence of kidney stone. After stone recurrence, 13 lithotomy surgeries were performed, and the SFR by the latest follow-up was 63.6%. CONCLUSION: This study had the longest follow-up time for patients with solitary kidney stones after PCNL reported at home and abroad. Ultrasound-guided standard PCNL was safe and effective in the treatment of solitary kidney stones. Long-term follow-up results showed that the recurrence rate of kidney stones was still high, but the long-term renal function was stable after operation, and some patients showed mild renal function decline.

[Clinical and pathological analysis of small renal cell carcinoma].

OBJECTIVE: To analyze the clinical and pathological features of small renal cell carcinoma (RCC), especially of those with diameter less than 4 cm and to understand the characteristics and factors related to recurrence and progression. METHODS: A total of 200 patients with RCC were stratifiedly selected for retrospective analysis. Their baseline demographic features, tumor-specific clinical features, pathological features of renal lesions, especially microscopic features were collected. The patients were divided according to the largest diameter of renal tumor lesions. Univariate analysis was used to compare the differences between tumor staging and microscopic pathological features between the groups. Binary multivariate Logistic regression was used to investigate factors related to tumor progression and prognosis in the patients with small RCC. RESULTS: The tumor diameters of 127 RCC patients were less than 4 cm and most of them had clear cell renal cell carcinoma (ccRCC). The increase in tumor diameter resulted in significantly higher T stage (P<0.01), higher WHO/International Society of Urological Pathology (ISUP) grade (P<0.05) and increasing chance of lymph node metastasis (P<0.01). Even when the tumor diameter was less than 4 cm, the patients might still have perirenal fat invasion, renal sinus invasion and greater elevated tumor grade (greater than grade 3) and synchronous lung metastasis. The incidences of intravascular thrombus (9.3% vs. 0) and tumor necrosis (27.8% vs. 5.5%) in the patients with RCC between 4-7 cm were significantly higher than those with RCC less than 4 cm (P<0.01). Sub-group analysis of small RCC (less than 4 cm) indicated that the patients with RCC between 2-4 cm were more likely to have intratumoral hemorrhage (44.7% vs. 23%, P<0.05) and necrosis than those with RCC less than 2 cm (8.2% vs. 0, P=0.095). Logistic regression analysis of small RCC showed that the incidence of tumor invasion to renal capsule was higher in ccRCC (OR=5.15, 95%CI: 1.36-19.52). Necrosis was closely related to the formation of peritumor pseudocapsule in small RCC (OR=14.90, 95%CI: 1.41-157.50). Increase in the tumor diameter was related to higher tumor grade (greater than grade 3) (OR=3.49, 95%CI: 1.11- 10.93). CONCLUSION: The tumor stage and grade of small RCC (less than 4 cm) are low, but extra-renal invasion and synchronous distant metastasis may occur. Internal hemorrhage and necrosis in tumor, ccRCC subtype, along with microscopic features, such as the renal capsule invasion and perirenal pseudocapsule formation are relevant factors of malignant behavior of small RCC and could be considered in prognosis evaluation.

[Altered serum cytokine expression profile in systemic sclerosis and its regulatory mechanisms].

OBJECTIVE: To analyze the expression profile of serum cytokines in patients with systemic sclerosis (SSc) and explore its possible regulatory mechanisms. METHODS: Serum and DNA of peripheral blood mononuclear cells were collected from 30 SSc patients and 80 normal controls (NCs). According to the presence or absence of interstitial lung disease (ILD) in SSc, the patients were divided into SSc with ILD group and SSc without ILD group. According to the degree of skin involvement, the patients were divided into diffuse systemic scleroderma (dcSSc) group and limited systemic scleroderma (lcSSc) group. According to the presence of anti-topoisomerase-1 antibody (anti-Scl-70 antibody) in the serum of patients with SSc, they were divided into SSc Scl-70 (+) group and SSc Scl-70 (-) group. 27 cytokines in serum were detected by Luminex MAGPIX detection system and Bio-Plex Pro Human Cytokine 27-plex Assay kit: interleukin-1ß (IL-1ß), interleukin-1 receptor antagonist (IL-1ra), IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12P70, IL-13, IL-15, IL-17, basic fiber growth factor (BASIC FGF), eotaxin, granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF), interferon-γ (IFN-γ), interferon-gamma induced protein 10(IP-10), monocyte chemotactic protein 1(MCP-1), macrophage inflammatory protein-1α (MIP-1α), macrophage inflammatory protein 1ß(MIP-1ß), platelet-derived growth factor BB (PDGF-BB), regulated on activation in normal T-cell expressed and secreted (RANTES), tumor necrosis factor-α (TNF-α), and vascular endothelial growth factor(VEGF). Methylation sites were detected by Illumina 450K methylation chip. RESULTS: Compared with NCs group, the expression of 12 cytokines (BASIC FGF, eotaxin, G-CSF, GM-CSF, IFN-γ, IL-1ß, IL-1ra, IL-6, IP-10, MCP-1, TNF-α and RANTES) in the SSc group significantly increased (P<0.05), IL-5 was decreased expression in the SSc group (P<0.05), there was no significant difference in the expressions of the other 14 cytokines. Compared with lcSSc group, 9 cytokines (eotaxin, IL-5, MCP-1, IL-2, RANTES, IL17A, IL-8, MIP-1ß and PDGF-BB) increased in dcSSc group, but there was no significant difference. Compared with SSc without ILD group, IL-15 increased in SSC with ILD group [18.2 (172.97) ng/L vs. 2.03(0.05) ng/L, P<0.05]. Compared with SSc Scl-70 (-) group, the expression of IP-10 decreased in SSc Scl-70 (+) group [1 030 (2 196.6) ng/L vs. 1 878 (2 964) ng/L, P<0.05]. The correlation analysis of serum cytokines with erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) showed that IL-6 was positively correlated with ESR (r =0.04, P= 0.017), MCP-1 (r=0.49, P=0.043) and MIP-1ß (r=0.41, P=0.007) positively correlated with CRP. By analyzing the changes of methylation sites of cytokines, it was found that cg17744604 in IL-10 TSS1500 region, cg06111286 in IL-12P70 TSS200 region, cg07935264 in IL-1ß TSS200 region, cg01467417 in IL-1ra TSS1500 region, cg03989987 in IL-1ra 5'UTR region and cg21099624 in VEGF TSS200 region were all hypomethylated. CONCLUSION: There were different cytokines expression profiles in the serum of SSc patients, and the altered cytokines were correlected with the degree of skin damage and pulmonary fibrosis. Many cytokines were regulated by methylation.

[A case of gastric duplication presenting as a retroperitoneal cystic lesion].

Gastric duplication is a rare congenital anomaly accounting to 4% of all gastrointestinal tract duplications. It is usually diagnosed pre-natally or during infancy. Retroperitoneal gastric duplication is very rare in adult patients. It is generally asymptomatic or presents with non-specific symptoms. The sensitivity of classical imaging modalities for retroperitoneal gastric duplication is weak. Retroperitoneal gastric duplication could be misdiagnosed as other retroperitoneal cystic lesions and it could be definitely diagnosed by typical findings of abdominal enhanced computed tomography (CT) and endoscopic ultrasonography (EUS) plus EUS:guided fine needle aspiration biopsy in gastric duplicated cyst. A female adult patient was diagnosed by an exceptional hospital CT as retroperitoneal cystic lesion and was admitted in our hospital in March 2019. The abdominal enhanced CT in our hospital preoperatively indicated a cystic lesion above the left kidney and the maximum cross section was 9.0 cm×5.2 cm. Lymphocysts and others should be identified and the lesion might be from the stomach. The retroperitoneal gastric duplication was not diagnosed before operation. In the operation laparoscopy revealed the cystic lesion in the retroperitoneum. The pancreas, left adrenal gland, and left kidney were compressed by the cystic lesion. The top of the lesion was connected with the small curvature of the stomach, but it was not communicated with the stomach. The retroperitoneal cystic lesion was completely excised and the surrounding organs were not damaged. The retroperitoneal gastric duplication was clearly diagnosed by the findings in the operation and the pathology of the lesion. The patient was discharged a week after surgery without gastrointestinal and pancreatic injuries. Therefore, the differential diagnosis of retroperitoneal cysts should consider the possibility of retroperitoneal gastric duplication. For patients with typical CT findings or unclear boundaries between the tumor and the stomach wall, gastric duplication could be diagnosed by EUS plus EUS:guided fine needle aspiration biopsy in the cyst. Retroperitoneal gastric duplication cyst could be completely excised by laparoscopy through abdominal cavity and the stomach wall could be completely repaired.

[Advances in epigenetic markers of dermatomyositis/polymyositis].

Idiopathic inflammatory myopathy (IIM) is a rare group of autoimmune diseases, characterized by chronic muscle weakness, muscle fatigue and infiltration of single nuclear cells in skeletal muscle. Its subtypes include dermatomyositis (DM), polymyositis (PM), inclusion body myositis (IBM) and immune-mediated necrotizing myositis (IMNM), and the most common subtypes are DM and PM. PM is an autoimmune disease mainly manifested by muscle damage. When the skin is involved, it is called DM. The incidence of IIM was relatively low, which was 1.16-19 per million people/year, but the mortality was high and the prognosis was poor. The pathogenesis of IIM is still unclear. Previous studies suggest that both immune and non-immune mechanisms are involved in its pathogenesis, especially cellular and humoral immunity. In recent years, researchers have conducted a number of studies on the pathogenesis of IIM, especially in the study of DM/PM with the application of high-throughput biometrics. Epigenetics is a discipline that refers to the genetic phenomena of DNA methylation spectrum, chromatin structure state and gene expression spectrum transferred between cells without any changes in DNA sequence, including DNA methylation, chromatin modification and non-coding RNA changes. A large number of studies have shown that epigenetic modification plays an important role in many diseases, especially in cancer. Recent studies have also found a series of epigenetic markers related to the occurrence and development of DM/PM, mainly in the aspect of non-coding RNA changes, such as miR-10a, miR-206, etc. And there has also been some research on DNA methylation. However, no studies have been reported on whether chromatin modification is involved in the pathogenesis of DM/PM. The pathogenesis of DM/PM is complex and diverse. With the development of research, certain microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) may become biological markers for the early diagnosis of DM/PM. Therefore, this paper mainly expounds the research progress of the biomarkers of DM/PM from the aspect of epigenetics.

[Clinical and pathological analysis of renal cell carcinoma bone metastasis].

OBJECTIVE: To analyze the clinical and pathological characteristics of renal cell carcinoma bone metastasis (RCC-BM) patients. METHODS: Data of RCC-BM patients from July 2003 to November 2017 were retrospectively reviewed. The patients' baseline characteristics (age, gender), tumor characteristics [specific sites of bone metastasis, time to bone metastasis (TTBM), imaging features of bone disease, coexistence of other metastasis], as well as pathological features (histological classification of primary and bone metastasis, immunohistochemical stain results) were collected. Descriptive analysis and difference analysis were used. RESULTS: A total of 113 RCC-BM patients were enrolled with the gender ratio (male:female) of 4:1, mean age of 59.39 years, and all present of osteolysis bone lesions. The common sites of bone metastasis were vertebra (46.0%) and pelvis (38.9%). Other distant metastasis sites coexisted in 28.3%, while 48.18% RCC-BM patients presented with synchronous metastasis (TTBM=0). The median TTBM for metachronous metastasis was 48 months. The majority in this cohort were determined to have primary tumor of clear cell carcinoma. After immunohistochemical examination to 104 RCC-BM patients and sub-group analysis, tendencies of higher positive rates of vascular endothelial growth factor (VEGF) was also found in synchronous group (P=0.097) while tendencies of higher positive rates of carbonic anhydrase (CA)-IX was found in the same group (P=0.100). The patients with clear cell RCC-BM had a significantly higher positive expression of epithelial growth factor receptor (EGFR, P<0.05) than those with non-clear cell RCC-BM group. CONCLUSION: More male and younger patients with metastatic lesions in axial skeleton were found in this cohort. Tendencies in the expression of CA-IX and VEGF in different TTBM sub-group and EGFR in different histology-derived subgroup indicate that they might be associated with risk and prognostic factors and support further target therapies of RCC-BM.