OBJECTIVE To investigate the relationship of long non-coding RNA （LncRNA） metastasis-associated lung adenocarcinoma transcript 1 （MALAT1） and doxorubicin （DOX） resistance in osteosarcoma （OS） cells. METHODS MG-63 and MG-63/DOX cells were treated with different concentrations of DOX （0， 0.01， 0.05， 0.1， 1 μmol/L）， and survival rates and half maximal inhibitory concentration were determined using CCK-8 assay. The expressions of LncRNA MALAT1 in MG-63 and MG-63/ DOX cells were detected by real-time quantitative fluorescence PCR. MG-63/DOX cells were divided into Control group， knocking down LncRNA MALAT1 negative control （sh-NC） group， sh-MALAT1 group， sh-MALAT1+anti-NC group， and sh-MALAT1+ anti-miR-154-5p group. The expressions of LncRNA MALAT1， miR-154-5p and cyclin D1 （CCND1） mRNA in MG-63/DOX cells of each group were detected. The effects of knocking down LncRNA MALAT1 on the proliferation， migration， invasion， and apoptosis of MG-63/DOX cells were detected by CCK-8 assay， scratch test， Transwell experiment and flow cytometry， respectively. The expression of proliferating cell nuclear protein （PCNA） and CCND1 protein in MG-63/DOX cells was detected by Western blot assay. Interactions between LncRNA MALAT1 and miR-154-5p， miR-154-5p and CCND1 were detected by dual luciferase reporter gene experiment. RESULTS Compared with 0 μmol/L DOX， 0.01， 0.05， 0.1 and 1 μmol/L DOX could reduce the survival rates of MG-63 and MG-63/DOX cells （except for 0.01 μmol/L DOX） （P＜0.05）， IC50 were 0.07 and 0.13 μmol/L， respectively. The survival rate， cell migration number and invasion number of MG-63/DOX cells， scratch closure rate， mRNA expressions of LncRNA MALAT1， mRNA and protein expressions of CCND1， and PCNA protein expression in sh-MALAT1 group were significantly lower than sh-NC group and Control group； the apoptosis rate and miR-154-5p expression were significantly higher than sh-NC group and Control group （P＜0.05）. sh-MALAT1+anti-miR-154-5p group was able to reverse the aforementioned biological effects in sh-MALAT1 group （P＜0.05）. In MG-63/DOX cells transfected with both MALAT1-wild type （WT） and CCND1-WT， the luciferase activity in the miR-154-5p mimic group was significantly lower than mimic negative control group （P＜ 0.05）. CONCLUSIONS Knocking down LncRNA MALAT1 can inhibit the DOX resistance of OS cells， and its mechanism may be targeting the miR-154-5p/CCND1 axis.

Objective: To establish a progressive research strategy for “colonic components analysis - efficacy verification and mechanism exploration - gut microbiota”, screen pharmacodynamic substances, and investigate their mechanism via gut microbiota. Methods: The pharmacodynamics of Gegen Qinlian decoction (GQD) were assessed using a mouse model of dextran sulfate sodium-induced ulcerative colitis (UC). Ultra-performance liquid chromatography-quadrupole-orbitrap mass spectrometer was used to identify the prototype and metabolic components of GQD in the colon during UC. To analyze the structure and function of characteristic genera of GQD and its active components, 16S rRNA sequencing was performed. Results: We identified 67 prototypic and 14 metabolic components of GQD in the UC colon. The primary prototype components are flavonoids and alkaloids, including puerarin (PUE), baicalin (BAI), and berberine (BER). The metabolism was predominantly sulfonation. Efficacy verification showed that the main active components, puerarin, baicalin, and berberine, had good therapeutic effects on UC. The results of 16S rRNA gene sequencing showed that GQD improved UC by regulating the structure and function of the gut microbiota. The abundance of gut microbiota involved in the metabolism of the prototype components was influenced by the corresponding components. The function prediction results showed that PUE was the most comparable to GQD, with 24 consistent pathways. BAI and BER showed comparable gut microbiota regulation pathways. Characteristic pathways of BER include glucometabolic processes. Conclusion This study focused on the key issues in the gut microbiota pathway and developed a progressive research strategy to understand the transformation mechanisms of colonic components. This research systematically analyzed the active components and metabolic transformation of GQD in the colon during the pathological state of UC, as well as changes in the structure and function of the gut microbiota, clarified the mechanism of GQD and its active components in improving UC via the gut microbiota pathway.

Objective:To analyze the status of anemia at the beginning of dialysis in maintenance hemodialysis (MHD) adult patients, and to explore the relationship between early dialysis anemia and early survival and long-term survival.Methods:It was a retrospective cohort study. The baseline demographic and clinical data of newly admitted MHD patients from January 1, 2013 to December 31, 2020 were retrospectively analyzed. According to the hemoglobin (Hb) level at the beginning of dialysis, the patients were divided into high Hb group (Hb≥110 g/L), middle Hb group (80 g/L≤Hb<110 g/L) and low Hb group (Hb<80 g/L). The baseline data among the three groups were compared, and the changing trend of Hb level in MHD patients during the 8 years was analyzed. The follow-up ended at peritoneal dialysis, kidney transplantation, death or on December 31, 2021. All-cause death event within 6 months after the initiation of dialysis was defined as early death, while all-cause death event more than 6 months after the initiation of dialysis was defined as long-term death. Kaplan-Meier survival curve was used to analyze the survival rate, and log-rank test was used to compare the survival rates among the three groups. Multivariate Cox regression analysis model was used to analyze the association between anemia (Hb<110 g/L) at the beginning of dialysis and both early and long-term mortality.Results:A total of 36 216 MHD patients were included in this study, with age of (61.3±15.5) years old and 22 163 males (61.20%). The Hb at the beginning of dialysis was (89.33±20.89) g/L. The compliance rate of Hb (≥110 g/L) was 16.43% (5 952/36 216). There were 12 232 patients (33.78%), 18 032 patients (49.79%), and 5 952 patients (16.43%) in low Hb group, middle Hb group, and high Hb group, respectively. There were statistically significant differences in gender distribution, age, serum creatinine, blood phosphorus, blood calcium, C-reactive protein, intact parathyroid hormone, blood leukocytes, platelets, serum albumin, triglyceride, total cholesterol, and proportions of chronic glomerulonephritis, diabetic nephropathy, diabetes mellitus, cardiovascular and cerebrovascular diseases, tumors, emporary catheter, long-term catheter and autologous arteriovenous fistula among the three groups (all P<0.05). During the 8-year period, the Hb level had an increased trend steadily each year, and Hb was (88.48±22.07) g/L, (88.52±21.43) g/L, (87.86±21.29) g/L, (88.93±20.69) g/L, (88.87±20.69) g/L, (90.03±20.47) g/L, (90.74±20.31) g/L and (90.31±20.54) g/L year by year. There were 2 176 early deaths (6.01%), and 6 557 long-term deaths (18.10%) by the end of follow-up. Kaplan-Meier survival curve showed that early survival rate of low Hb group was significantly lower than those of high Hb group (log-rank test, χ2=57.115, P<0.001) and middle Hb group (log-rank test, χ2=49.918, P<0.001), and long-term survival rates of low Hb group (log-rank test, χ2=107.097, P<0.001) and middle Hb group (log-rank test, χ2=47.430, P<0.001) were significantly lower than that of high Hb group. Multivariate Cox regression analysis showed that Hb<80 g/L at the beginning of dialysis was an independent influencing factor of early death (Hb ≥110 g/L as a reference, HR=1.307, 95% CI 1.096-1.559), and 80 g/L≤Hb<110 g/L and Hb<80 g/L at the beginning of dialysis were the independent influencing factors of long-term death (Hb≥110 g/L as a reference, HR=1.108, 95% CI 1.021-1.203; HR=1.228, 95% CI 1.127-1.339, respectively) in MHD patients. Conclusions:The compliance rate of Hb at the beginning of dialysis in MHD patients is low. Hb <80 g/L at the beginning of dialysis is an independent risk factor of early death, and Hb <110 g/L at the beginning of dialysis is an independent risk factor of long-term death in MHD patients.

Objective:To determine the impact of early serum sodium concentrations on the survival prognosis in maintenance hemodialysis (MHD) patients.Methods:It was a retrospective cohort study. The newly admitted hemodialysis patients who were included in the registration system of Zhejiang Province Dialysis Quality Control Center from January 1, 2010 to December 31, 2019 were identified. Follow-up was conducted until December 31, 2020. Baseline data were collected for the first three months of dialysis, in which the mean level of serum sodium was defined as early serum sodium. Patients were divided into five groups based on early serum sodium level. Restricted cubic spline (RCS) was used to fit the relationship between long-term serum sodium level and risk of death. Kaplan-Meier model and Log-rank test were used to compare the survival rates of different groups. Multivariable Cox regression was used to analyze the correlation between early serum sodium level and death.Results:A total of 26 309 MHD patients were included in this study, and their ages were (59.07±15.41) years (ranging from 18 to 100 years). Among them, 13 643 (51.9%) were over 60 years old and 15 843 (60.2%) were males. Among the primary diseases of chronic renal failure, chronic glomerulonephritis was the first [13 703 cases (52.1%)], followed by diabetic nephropathy [6 460 cases (24.6%)], hypertensive nephropathy [1 293 cases (4.9%)], polycystic kidney disease [1 164 cases (4.4%)], etc. According to early serum sodium level, 12 883 patients (49.0%) had hyponatremia (serum sodium <135 mmol/L), of which 4 001 patients (15.2%) had serum sodium ≤130 mmol/L; 1 529 patients (5.8%) had hypernatremia (serum sodium >145 mmol/L). Patients were divided into the following 5 groups: 4 001 cases (15.2%) in group 1 (serum sodium ≤130 mmol/L), 8 882 cases (33.8%) in group 2 (130 145 mmol/L). Among them, patients in the Na≤130 mmol/L group had a slightly older age, a higher proportion of diabetes and cardiovascular disease, a lower level of blood uric acid, albumin, hemoglobin, and a higher level of alkaline phosphatase and leukocytes, while patients in the Na >145 mmol/L group had an older age and a higher proportion of cardiovascular disease. After follow-up of (55.67±33.58) months, a total of 4 954 patients (18.8%) died, 1 537 patients (5.8%) underwent kidney transplantation, 128 patients (0.5%) were converted to peritoneal dialysis. Of the deaths, 990 (20.0%) were due to cardiovascular diseases, 498 (10.1%) to cerebrovascular diseases and 400 (8.1%) to infections, and cardiovascular disease was the main cause of death. RCS curve fitting of the relationship between serum sodium level and risk of death found that the all-cause mortality hazard ratio ( HR) increased with decreasing or increasing serum sodium, and the optimal serum sodium was between 135 mmol/L and 140 mmol/L. Kaplan-Meier survival curve showed that the risk of all-cause death (Log-rank test, χ2=66.5, P<0.001), the risk of cardiovascular death (Log-rank test, χ2=31.5, P<0.001) and the risk of infection death (Log-rank test, χ 2=28.6, P<0.001) were significantly different among five groups. The 10-year cumulative survival rate was 63.0%, 71.5%, 72.5%, 67.8% and 61.4% in groups with different serum sodium levels from low to high, and the 10-year cumulative cardiovascular mortality rate was 9.6%, 6.2%, 5.5%, 7.3% and 11.7%, and the 10-year cumulative infection mortality rate was 4.9%, 3.2%, 1.7%, 2.8% and 3.9%. Multivariable Cox regression showed early serum sodium level >145 mmol/L was an independent relevant factor for all-cause death in MHD patients ( HR=1.237, 95% CI 1.045-1.465, P=0.013). Conclusions:MDH patients are more likely to develop hyponatremia in the early stage of dialysis. The cumulative survival rate of all-cause death, cardiovascular death and infection death in patients with predialysis serum sodium ≤130 mmol/L and >145 mmol/L within three months after initiation of dialysis is significantly lower than those in other levels. Early serum sodium >145 mmol/L is associated with higher mortality in MHD patients.

Objective:To study the relationship and the role of leptin in children with biliary atresia and hepatic fibrosis to provide a treatment basis for these patients.Methods:The clinical data of children with biliary atresia or congenital biliary dilatation (CBD) who underwent surgical treatment at the Department of General Surgery of Tianjin Children's Hospital from August 2019 to August 2021 were retrospectively analyzed. Of 31 children included in this study, there were 14 males and 17 females, with age of 60 (30, 63) d. Children with biliary atresia served as the study group ( n=26) and children with CBD served as the control group ( n=5). Leptin protein, α-smooth muscleactin (α-SMA) and phosphorylation of extracellular-regulated protein kinase 1/2 (p-ERK1/2) in liver tissues were detectd by immunohistochemistry (IHC). The expression level of leptin mRNA in liver tissues were detected by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Results:The average optical density values of leptin protein, α-SMA protein and p-ERK1/2 protein in the liver tissues of children in the study group were significantly higher than the control group ( P<0.05). The expression levels of leptin, α-SMA and p-ERK1/2 in liver tissues of children with biliary atresia significantly increased with increase in fibrosis degree ( P<0.05). The expression level of leptin in liver tissues of children with biliary atresia was positively correlated with the liver fibrosis grade ( rs=0.876), α-SMA ( r=0.723) and p-ERK1/2 ( r=0.725) ( P<0.01). The results of qRT-PCR showed that the content of leptin mRNA in liver tissues of children with biliary atresia was significantly higher than that of children with CBD ( P<0.05). Conclusion:Expressions of leptin increased with aggravation of degrees of hepatic fibrosis in biliary atresia. Leptin may be involved in activation of HSCs through the ERK1/2 signaling pathway in the process of hepatic fibrosis due to biliary atresia.

BACKGROUND: The expression of programmed cell death ligand 1 (PD-L1) as a biomarker for immunotherapy in non-small cell lung cancer (NSCLC) is routinely detected in clinical pathology department. However, the spatial heterogeneity of PD-L1 expression in intrapulmonary tumors and extrapulmonary metastases is still a challenge for the clinical testing. This study aims to explore the differences of PD-L1 expression in test samples obtaining from different sites of NSCLC. This study may contribute to the detection strategy of PD-L1 in patients with advanced lung cancer. METHODS: One hundred and thirty-one cases of consecutively detected PD-L1 (22c3 assay, Dako) staining in metastatic NSCLC and 972 cases of non-paired intrapulmonary NSCLC were collected. The discrepancies of tumor proportion score (TPS) of PD-L1 expression in intrapulmonary samples and extrapulmonary metastatic samples of different sites were compared. RESULTS: The positive expression rate of PD-L1 in extrapulmonary metastatic NSCLC (TPS ≥ 1%) was 61.83%, and the TPS was significantly higher than that in intrapulmonary tumors (P=0.03). The PD-L1 scores of the specimens obtained from different sites were significantly different (P=0.007). The positive rates of PD-L1 in liver and adrenal metastases were 85.71% and 77.78% respectively, and their TPS were significantly higher than that of the intrapulmonary samples (P<0.05). The positive rates of PD-L1 in lymph node, bone, brain, soft tissue, and pleural metastases was 40.00%-66.67%, with no significant differences compared to intrapulmonary tumors. The analysis of histological subtype and sample type showed that the PD-L1 score of extrapulmonary samples of adenocarcinoma subtype or surgical specimen was significantly higher than that of intrapulmonary tumors. The analysis of clinicopathological parameters showed that the PD-L1 positive expression or high expression were significantly correlated with male patients, smoking history, and epidermal growth factor receptor (EGFR) wild type. CONCLUSIONS The expression of PD-L1 in metastatic NSCLC is generally higher than that in intrapulmonary tumor, and the positive rate of PD-L1 expression was discrepant in different sites of specimen. The differences of PD-L1 score between extrapulmonary metastatic samples and intrapulmonary samples may be associated with different metastatic sites, histological subtype, and specimen type.
B7-H1 Antigen/metabolism* ; Biomarkers, Tumor/metabolism* ; Carcinoma, Non-Small-Cell Lung/drug therapy* ; Humans ; Immunohistochemistry ; Lung Neoplasms/drug therapy* ; Male

Objective:To retrospectively analyze the early mortality and related risk factors in adult patients with maintenance hemodialysis (MHD).Methods:Adult MHD patients from 2008 to 2018 were enrolled and divided into training data group and validation data group. In training data group, multivariate logistic regression was used to analyze the risk factors of early death within 120 days after hemodialysis and establish a prediction model. The receiver operating characteristic (ROC) curve was applied to evaluate the prediction ability of the model.Results:A total of 4 885 patients were included. The cumulative mortality within 120 days was 20.97/100 person years, and that within 365 days was 12.25/100 person years. A total of 3 603 patients in the training data group were analyzed. The following risk factors were correlated with early mortality (all P<0.05), including age at start of dialysis over 60 years old ( OR=1.792), non-chronic glomerulonephritis ( OR=2.214), cardio-cerebrovascular disease ( OR=2.695), plasma albumin less than 35 g/L ( OR=1.358), platelet count less than 120×10 9/L ( OR=2.194), serum creatinine less than 600 μmol/L ( OR=1.652), blood urea nitrogen over 30 mmol/L ( OR=1.887), blood phosphorus less than 1.13 mmol/L ( OR=1.783), pulse pressure over 55 mmHg(1 mmHg=0.133 kPa) ( OR=1.656), low density lipoprotein less than 1.5 mmol/L ( OR=1.873), and blood calcium over 2.5 mmol/L ( OR=1.876). Risk prediction model was established. The other 1 282 cases in the validation data group were verified. The area under ROC curve was 0.810, with sensitivity 85.7%, and specificity 62.5%. Conclusion:The mortality rate of adult MHD patients within 120 days after dialysis is high. The established prediction model can effectively predict the risk of early death.

Objective:To explore the relationship between end-dialysis over-weight (edOW) in initial stage of hemodialysis and long-term prognosis in maintenance hemodialysis patients.Methods:The data of initial uremia patients receiving hemodialysis in the First Affiliated Hospital, College of Medicine, Zhejiang University from January 1, 2008 to April 30, 2017 were retrospectively analyzed. The end point of follow-up was death or until April 30, 2018. The general data including age, gender, body mass index, primary disease, complications and laboratory indicators of the patients and the related parameters of dialysis from four to twelve months were collected. Kaplan-Meier method was used to analyze survival rate. Cox multivariate regression was used to analyze the relationship between edOW and all-cause mortality and cardiovascular disease (CVD) mortality.Results:A total of 469 patients (300 males, 64.0%) were enrolled, with age of (56.9±17.1)years old. During the follow-up period of (4.1±2.4) years (1.0-10.3 years), 102 patients died. The main cause of death was cardiovascular and cerebrovascular events, accounting for 44.1% (45/102). The value of edOW was (0.28±0.02) kg. The patients were divided into edOW<0.28 kg group ( n=292) and edOW≥0.28 kg group ( n=177) according to the mean value of edOW. Kaplan-Meier survival analysis showed that the long-term survival rate in edOW<0.28 kg group was higher than that in edOW≥0.28 kg group (Log-rank χ2=4.134, P=0.043), and the CVD mortality in edOW≥0.28 kg group was significantly higher than that in edOW<0.28 kg group (Log-rank χ2=11.136, P=0.001). Cox multivariate regression analysis showed that high edOW was an independent influencing factor for all-cause death and CVD death in hemodialysis patients ( HR=1.541, 95% CI 1.057-2.249, P=0.025; HR=1.930, 95% CI 1.198-3.107, P=0.007). Conclusion:High edOW in early phase is an independent influencing factor of all-cause and CVD death in hemodialysis patients.

During pandemic of corona virus disease 2019 (COVID-19), emergency orthopedic trauma is commonly seen. It is particularly important to ensure the emergency treatment quality of orthopedic trauma but avoid cross-infection between doctors and patients. The double-buffered diagnosis and treatment mode refers to the model of patients first undergoing medical observation in the comprehensive buffer ward and the inpatient buffer rooms of various disciplines after admission to confirm the exclusion of COVID-19 and then receiving specialist diagnosis and treatment. The authors summarize the experiences of using the double-buffered diagnosis and treatment model in the Department of Orthopedics, Renmin Hospital of Wuhan University during the prevention and control of COVID-19 pandemic so as to provide a reference for treatment of orthopedic patients.

Objective: To explore the relationship between serum C1q and tumor necrosis factor related protein 6(CTRP6) level and insulin resistance in patients with newly diagnosed type 2 diabetes mellitus (T2DM). Methods: A total of 167 patients with newly diagnosed T2DM in the outpatient department of our hospital were recruited from April 2016 to March 2017 and 165 subjects with normal glucose tolerance were used as the control group. The concentrations of CTRP6, interleukin 6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor α (TNF-α) were determined by ELISA. Results: Circulating CTRP6 level was significantly higher in T2DM group than that in control group [(652.54±132.57) vs (521.28±119.93)μg/L, P<0.01] after adjusting age and body mass index (BMI). Overweight/obese subjects revealed higher CTRP6 levels compared with those in lean individuals. In addition, circulating CTRP6 level was positively correlated with BMI, waist circumference, fasting plasma glucose, postprandial 2h plasma glucose, HbA_1C, fasting insulin, homeostasis model assessment insulin resistance index (HOMA-IR), triglyceride (TG), IL-6, MCP-1, highly sensitive C-reactive protein (hs-CRP), and TNF-α, while it was inversely correlated with high-density lipoprotein-cholesterol(P<0.01). Multivariate linear regression analysis showed that TG, HOMA-IR, and IL-6 were independent factors for CTRP6 level. After adjusting for potential confounders, CTRP6 remained an independent risk factor for T2DM. Trend test showed that the increase in CTRP6 level was significantly linear with the occurrence of T2DM. The analysis of receiver operating characteristic curves revealed that the area under the curve for circulating CTRP6 to predict T2DM was 0.730. Conclusions CTRP6 may be associated with insulin resistance.