Objective To investigate the epidemic characteristics and factors influencing mortality in human immunodeficiency virus (HIV) and hepatitis C virus (HCV)-coinfected patients in heilongjiang region. Methods The clinical data of 712 HIV-infected patients attending our hospital from January 2019 to December 2023 were analyzed retrospectively. The HCV infections were detected by ELISA, and epidemic characteristics of HIV/HCV co-infected patients were analyzed . Co-infected patients (n=116) were classified into survival group (n=90) and death group (n=26) according to the follow-up results, and risk factors for the mortality of HIV/HCV co-infected patients were identified. Results The prevalence of HIV/HCV co-infection was 16.29%, and the prevalence was the highest in 30-50 age group, accounting for 25.47%. There was no significant difference in co-infection rates by gender and marital status (P>0.05). The co-infection rate was the highest among farmers (26.85%) by occupation and among intravenous drug users (61.90%) by route of infection. No statistical difference was found in gender, marital status, and occupational status between survival group and death group (P>0.05), whereas statistical difference was found in age at diagnosis of HIV infection, route of infection, antiretroviral treatment, percentage of CD4+T lymphocytes, and interval between diagnosis and initiation of treatment between two groups (P<0.05). Multivariate Logistic regression analysis denoted that age at diagnosis of HIV infection (OR=1.827, 95% CI: 1.263-2.722), route of infection (OR=1.796, 95% CI: 1.248-2.503), antiretroviral treatment (OR=1.724, 95% CI: 1.202-2.367), percentage of CD4+T lymphocytes (OR=2.536, 95% CI: 1.776-3.739), and the time interval between diagnosis and initiation of treatment (OR=1.953, 95% CI: 1.431-2.952) were all associated with the risk of mortality in co-infected patients (P<0.05). Conclusion The overall prevalence of HIV/HCV co-infection is 16.29% in heilongjiang region . In order to reduce the mortality rate of superinfection , we should focus on patients who are ≥ 40 years old, intravenous drug use , no antiviral therapy , low CD4+ T lymphocyte levels for the first time , and have a long interval between diagnosis and initiation of treatment.

Objective:To establish a nomogram model for predicting the risk of sepsis in diabetic foot patients, and to provide reference for clinical prevention and treatment.Methods:The clinical data of 430 patients with diabetic foot who were hospitalized in Chu Hsien-I Memorial Hospital of Tianjin Medical University from January 2022 to March 2023 were reviewed and collected, including age, gender, past medical history, smoking and drinking history, family history, diabetes course, Texas grade of diabetic foot and laboratory indicators within 24 hours after admission. Patients were divided into sepsis group and non-sepsis group according to the presence or absence of sepsis during hospitalization. The differences in clinical data between the two groups were compared. Multivariate Logistic regression analysis was used to screen the influencing factors of sepsis in patients with diabetic foot during hospitalization, and a nomogram predictive model was established. The performance of the prediction model was evaluated by receiver operator characteristic curve (ROC curve), calibration curve and decision curve analysis (DCA). Internal validation was performed by using Bootstrap method.Results:A total of 430 patients were enrolled, among which 90 patients developed sepsis during hospitalization and 340 patients did not. There were statistically significant differences in diabetes course, Texas grade of diabetic foot, white blood cell count (WBC), neutrophil count (NEU), lymphocyte count (LYM), neutrophil to lymphocyte ratio (NLR), hemoglobin (Hb), albumin (Alb), glycosylated hemoglobin (HbA1c), C-reactive protein (CRP), and blood urea nitrogen (BUN) between the two groups. Multivariate Logistic regression analysis showed that diabetes course [odds ratio ( OR) = 2.774, 95% confidence interval (95% CI) was 1.053-7.308, P = 0.039], Texas grade of diabetic foot ( OR = 2.312, 95% CI was 1.014-5.273, P = 0.046), WBC ( OR = 1.160, 95% CI was 1.042-1.291, P = 0.007), HbA1c ( OR = 1.510, 95% CI was 1.278-1.784, P < 0.001), CRP ( OR = 1.007, 95% CI was 1.000-1.014, P = 0.036) were independent risk factors for sepsis in patients with diabetic foot during hospitalization, while Alb was a protective factor ( OR = 0.885, 95% CI was 0.805-0.972, P = 0.011). A nomogram predictive model was constructed based on the above 6 indicators. The ROC curve showed that the area under ROC curve (AUC) of the nomogram predictive model for identifying the sepsis patients was 0.919 (95% CI was 0.889-0.948). The AUC of the nomogram predictive model after internal verification was 0.918 (95% CI was 0.887-0.946). Hosmer-Lemeshow test showed χ2 = 2.978, P = 0.936, indicating that the calibration degree of the predictive model was good. Calibration curve showed that the predicted probability of sepsis was in good agreement with the actual probability. DCA curve showed that the nomogram predictive model had good clinical usefulness. Conclusion:The nomogram predictive model based on the risk factors of diabetes course, Texas grade of diabetic foot, WBC, HbA1c, CRP and Alb has good predictive value for the occurrence of sepsis in patients with diabetic foot during hospitalization, which is helpful for clinical screening of the possibility of diabetic foot patients progressing to sepsis, and timely personalized intervention for different patients.

Objective:To evaluate the clinical application value of iohexol plasma clearance assay in assessing glomerular filtration rate (GFR) in patients with chronic kidney disease (CKD), and identify alternative methods of 99mTc-diethylene triamine pentaacetic acid renal kinetic imaging (Gates) method for measuring GFR ( 99mTc-mGFR). Methods:It was a cross-sectional study. The CKD patients hospitalized in the Department of Nephrology at the Second Affiliated Hospital of Nanjing Medical University between October 2022 and September 2023 were enrolled. Iohexol plasma clearance was determined by collecting blood samples at 2 and 4 hours after intravenous administration of 5 ml iohexol, and high performance liquid chromatography was used to measure the plasma concentration of iohexol. Br?chner-Mortensen and Jacobsson formulas were used to calculate the double and single plasma iohexol clearance, respectively. CKD-epidemiology collaboration equation based on serum creatinine concentration was used to calculate the estimated GFR (eGFR). Pearson's coefficient was performed to analyze the correlation of iohexol dual plasma clearance assay-measured GFR (iohexol-DS-mGFR), iohexol single plasma 4 hours clearance assay-measured GFR (iohexol-SS 4h-mGFR), iohexol single plasma 2 hours clearance assay-measured GFR (iohexol-SS 2h-mGFR) and eGFR with 99mTc-mGFR as the gold standard. Bland-Altman analysis, 95% limits of agreement, and intra-correlation coefficient were used to compare the diagnostic concordance of Iohexol-DS-mGFR, Iohexol-SS 4h-mGFR, Iohexol-SS 2h-mGFR and eGFR with 99mTc-mGFR. Results:The study enrolled 64 CKD patients, aged (58.91±13.08) years old, comprising of 38 males and 26 females. The distribution of patients across CKD stages based on 99Tc-mGFR was as follows: 12 patients (18.8%) in stage 1, 14 patients (21.9%) in stage 2, 26 patients (40.6%) in stage 3, 10 patients (15.6%) in stage 4, and 2 patients (3.1%) in stage 5. The Pearson correlation analysis revealed that the correlation coefficients of iohexol-DS-mGFR, iohexol-SS 4h-mGFR, iohexol-SS 2h-mGFR and eGFR with 99mTc-mGFR were 0.925, 0.867, 0.820 and 0.894 (all P<0.001), respectively. The median deviation of absolute value of iohexol-DS-mGFR, iohexol-SS 4h-mGFR, iohexol-SS 2h-mGFR and eGFR in the total study population were 6.66, 9.63, 11.47 and 9.59 ml·min -1·(1.73 m 2) -1, respectively. The proportions of iohexol-DS-mGFR, iohexol-SS 4h-mGFR, iohexol-SS 2h-mGFR and eGFR located in the gold-standard GFR ± 10% interval (P10) were 35.9%, 29.7%, 26.6% and 29.7%, respectively, and the proportions located in the gold-standard GFR ± 30% interval ( P30 ) were 87.5%, 68.8%, 60.9% and 73.4%, respectively. Conclusions:Iohexol dual plasma clearance is well correlated and consistent with 99mTc-mGFR. It is a safe and easy alternative to isotope 99mTc-mGFR for clinical use in determining GFR.

AIM:To investigate whether the expression of perilipin 2(PLIN2)in renal tubular cells could predict a decline in renal function in diabetic kidney disease(DKD)patients,and to explore the potential mechanisms in-volved in renal tubular cell injury induced by PLIN2 during the progression of DKD.METHODS:Control individuals(n=12)and DKD patients(n=51)were enrolled in this retrospective cohort study.Demographic and laboratory data were col-lected.A simplified linear mixed-effects model was applied to assess the estimated glomerular filtration rate(eGFR)slope.The relationship between PLIN2 and renal function decline in DKD patients was predicted by Spearman correlation analysis and a generalized linear model.BKS-db/db diabetic mice and streptozotocin-induced diabetic mice were used.Primary renal tubular cells were treated with glucose and transfected with small interfering RNA or plasmid.Western blot-ting and immunofluorescence staining were used to detect PLIN2 expression.Lipid droplets were stained with oil red O.The oxygen consumption rate(OCR)of mitochondria was measured using an extracellular flux analyser.RESULTS:The expression of PLIN2 was markedly higher in the tubules of DKD patients than in those of control subjects.After 24(12,39)months of follow-up,the eGFR slope of DKD patients was-7.42(-19.77,-2.09)mL/(min·1.73 m2·year).An in-crease in the baseline percentage of PLIN2-positive tubules was significantly associated with the eGFR slope during the fol-low-up period[hazard ratio(HR)=1.90,95%confidence interval(CI):1.00～3.58],indicating that tubular PLIN2 could predict a decrease in renal function in DKD patients.Both the accumulation of lipid droplets and the expression of PLIN2 were markedly greater in the tubules of diabetic mice than in those of control mice.Glucose treatment induced lipid droplet accumulation and PLIN2 expression in renal tubular cells.Knockdown of PLIN2 significantly alleviated glucose-in-duced lipid droplet accumulation,whereas PLIN2 overexpression aggravated glucose-induced lipid droplet accumulation.The decrease in mitochondrial OCR in renal tubular cells induced by glucose treatment was alleviated after PLIN2 knock-down.However,overexpression of PLIN2 directly decreased the mitochondrial OCR.CONCLUSION:The PLIN2 ex-pression in tubules predicts a decline in renal function in patients with DKD.The PLIN2 suppresses mitochondrial aerobic respiration and contributes to the accumulation of lipid droplets in renal tubular cells to promote the progression of DKD.

Objective To explore the mechanism of miR-421 affecting the occurrence and development of depression.Methods A depressive rat model was established by single intraperitoneal injection of lipopolysaccharide(LPS),and depressive behavior was detected by glucose preference test and open-field test.miRNA microarray chips and RT-PCR were used to analyze the expression level of miR-421 in hippocampus of the depressed rats.TargetScan database and mi RDB database were used to predict the target genes of miR-421.Dual luciferase reporter gene assay was used to observe the binding of miR-421 to the target genes.The impact of over-expression and inhibition of miR-421 on target genes was observed,then the influence of over-expression and inhibition of target genes on downstream factors was observed,and the related mechanism of miR-421 on depression was explored.Results miRNA microarray chips and RT-PCR assay showed that miR-421 was highly expressed in the hippocampus of the depressed rats(P<0.001),Inhibition of miR-421 expression could significantly restore the body weight and exercise ability of the depressed rats(P<0.001).Binding targets of Menin and miR-421 were predicted by TargetScan database,and interaction between Menin and miR-421 was demonstrated by dual-luciferase reporter gene assay.Menin expression was down-regulated while miR-421 was overexpressed(P<0.001),whereas it was up-regulated as miR-421 was inhibited(P<0.001).qPCR indicated that expressions of Caspase-3 and NF-κB in the hippocampus of the depressed rats was significantly increased(P<0.001),and IL-1β expression in the hippo-campus was significantly increased(P<0.01).When the expression of Menin was inhibited,the expressions of Caspase-3,NF-κB and IL-1β were increased(P<0.001),while the expressions of Caspase-3,NF-κB and IL-1β were decreased when Menin was overexpressed(P<0.001).Conclusions Inhibition of miR-421 expression can increase Menin expression,decrease Caspase-3 content,and reduce neuroinflammatory response,thereby improving depressive symptoms.

Objective To develop a suitable medium and optimize culture time for the primary osteoblast culture from suckling mouse,so to provide an improved experimental protocol for primary osteoblast culture in vitro.Meth-ods Primary osteoblasts were collected from skull of CD1 suckling mouse by interrupted enzyme digestion.The pu-rified osteoblasts were harvested by differential centrifugation.The incubation time,concentration of fetal bovine se-rum(FBS),β-glycerophosphate sodium and dexamethasone were tested and optimized.The change of osteoblast maturation marker was examined by Western blot(WB)and immunofluorescence staining(IF).The osteogenic ac-tivity was determined by alkaline phosphatase staining,alizarin red staining and ultrastructure.Results Primary osteoblast were obtained from sucleling mouse skull bone by interrupted enzyme digestion for proliferation and trans-generational expansion.The expression of osteoblast maturation markers was parallel to the time of induction culture and the concentration of FBS.Mature osteoblasts were obtained by culturing the cells with 10％ FBS for 14 days.The differentiation of primary osteoblasts was induced by different concentrations of β-glycerophosphate and dexam-ethasone.The results showed that the expression of osteoblast maturation markers was higher under the culture con-ditions of 10 mmol/L β-glycerophosphate and 5 nmol/L dexamethasone(P<0.01),and the staining of alkaline phosphatase and alizarin red was obvious,and the osteogenic activity was better too.Conclusions Primary osteo-blasts isolated from the skull of suckling CD1 mice cultured in induction medium containing 10％fetal bovine ser-um,10 mmol/L β-glycerophosphate sodium and 5 nmol/L dexamethasone for 14 days show good osteogenic activity and are suitable for in vitro experimental studies.

OBJECTIVE To explore the application value of indocyanine green combined with near-infrared autofluorescence imaging technique in identifying pathological parathyroid glands during surgery for primary hyperparathyroidism (PHPT). METHODS Data from 40 patients with PHPT treated in Beijing Tongren Hospital,Capital Medical University were collected,including 10 patients in the indocyanine green treated group and 30 patients in the non-treated group. All patients underwent surgical treatment to remove the affected parathyroid glands. Near-infrared autofluorescence imaging was used for image acquisition,and ImageJ software was used for fluorescence intensity analysis. RESULTS The fluorescence intensity of the pathological parathyroid glands in the indocyanine green-treated group was significantly higher than that in the non-treated group(142.7±23.7 vs. 94.5±31.4,t=-4.434,P=0.000);the fluorescence ratio of pathological parathyroid glands/thyroid glands was significantly higher than that in the non-treated group(1.6±0.3 vs. 1.2±0.4,t=-3.162,P=0.004). There was no correlation between the fluorescence intensity of parathyroid glands in the non-treated group and preoperative blood calcium(r=0.029,P=0.088) and preoperative PTH level(r=-0.142,P=0.455),and there was also no correlation between the fluorescence intensity of parathyroid glands in the treated group and preoperative blood calcium(r=0.206,P=0.568) and preoperative PTH level(r=0.160,P=0.658). The detection rate of near-infrared light in the non-treated group was 53.3％(16/30),while that in the treated group was 100％(10/10). The average detection time for the non-treated group was (71.0±16.9)minutes,while that for the treated group was (52.7±11.1)minutes,with a significant difference between the two groups(t=3.187,P=0.003). CONCLUSION The combination of indocyanine green and near-infrared autofluorescence imaging technique is helpful for identifying the diseased parathyroid glands during the surgical treatment of PHPT.

Background::Heterogeneity of tumor cells and the tumor microenvironment (TME) is significantly associated with clinical outcomes and treatment responses in patients with urothelial carcinoma (UC). Comprehensive profiling of the cellular diversity and interactions between malignant cells and TME may clarify the mechanisms underlying UC progression and guide the development of novel therapies. This study aimed to extend our understanding of intra-tumoral heterogeneity and the immunosuppressive TME in UC and provide basic support for the development of novel UC therapies.Methods::Seven patients with UC were included who underwent curative surgery at our hospital between July 2020 and October 2020. We performed single-cell RNA sequencing (scRNA-seq) analysis in seven tumors with six matched adjacent normal tissues and integrated the results with two public scRNA-seq datasets. The functional properties and intercellular interactions between single cells were characterized, and the results were validated using multiplex immunofluorescence staining, flow cytometry, and bulk transcriptomic datasets. All statistical analyses were performed using the R package with two-sided tests. Wilcoxon-rank test, log-rank test, one-way analysis of variance test, and Pearson correlation analysis were used properly.Results::Unsupervised t-distributed stochastic neighbor embedding clustering analysis identified ten main cellular subclusters in urothelial tissues. Of them, seven urothelial subtypes were noted, and malignant urothelial cells were characterized with enhanced cellular proliferation and reduced immunogenicity. CD8 + T cell subclusters exhibited enhanced cellular cytotoxicity activities along with increased exhaustion signature in UC tissues, and the recruitment of CD4 + T regulatory cells was also increased in tumor tissues. Regarding myeloid cells, coordinated reprogramming of infiltrated neutrophils, M2-type polarized macrophages, and LAMP3 + dendritic cells contribute to immunosuppressive TME in UC tissues. Tumor tissues demonstrated enhanced angiogenesis mediated by KDR + endothelial cells and RGS5 +/ACTA2 + pericytes. Through deconvolution analysis, we identified multiple cellular subtypes may influence the programmed death-ligand 1 (PD-L1) immunotherapy response in patients with UC. Conclusion::Our scRNA-seq analysis clarified intra-tumoral heterogeneity and delineated the pro-tumoral and immunosuppressive microenvironment in UC tissues, which may provide novel therapeutic targets.

Objective:To construct an evaluation index system for continuous nursing outcomes of patients undergoing knee arthroplasty.Methods:From January to May 2022, a preliminary draft of the index system was developed based on literature research, semi-structured interviews, and group discussions. We developed a questionnaire and conducted two rounds of consultation with 15 experts using the Delphi expert consultation method to construct the final index system. The positivity of experts was expressed by the effective recovery rate of the questionnaire. The expert authority coefficient was represented by the arithmetic mean of the expert's judgment basis and familiarity with the indicator. The coordination of expert opinions was represented by the Kendall's W. Results:Around two rounds of consultation, the effective recovery rates of the questionnaire were 100.0% (15/15) and 93.3% (14/15), respectively, and the expert authority coefficients were 0.815 and 0.860, respectively, and the expert Kendall's W were 0.297 and 0.121 ( P<0.01). The final evaluation index system for continuous nursing outcomes of patients undergoing knee arthroplasty included 3 primary indicators, 7 secondary indicators, and 47 tertiary indicators. Conclusions:The evaluation index system for continuous nursing outcomes of patients undergoing knee arthroplasty is strongly scientific and reliable, and has guiding significance for standardizing the continuous nursing of knee arthroplasty patients.

Objective: To study anti⁃hepatitis C virus NS3/4A protease activity of Lonicera fulvotomentosa based on fluorescence method and molecular docking. Methods: The activity of HCV NS3/4A protease inhibition of L.fulvotomentosa extracts was tested by fluorescence method. The binding of the main active ingredients to HCV NS3/4A virus protease was analyzed based on molecular docking. Results: The water extract and alcohol extract of L.fulvotomentosa had a good inhibitory effect on the activity of HCV NS3/4A protease with IC50 of 0. 005 - 0. 019 mg/ml.Based on molecular coupling and interaction analysis , it was found that chlorogenic acid , isochlorogenic acid A , cynaroside and luteolin combined well with HCV NS3/4A protease , forming multiple hydrogen bonds. Conclusion The active components of L. fulvotomentosa against HCV NS3/4A protease activity were isochlorogenic acid A and luteolin.