Objective: To explore the mechanism of action of Zangjiangzhi capsule (ZJZC) in treating hyperlipidemia (HLP). Methods: The components of ZJZC were analyzed and identified using ultra-high performance liquid chromatography with Q-Exactive Orbitrap tandem mass spectrometry (UHPLC-Q-Exactive-Orbitrap-MS/MS). Network pharmacology analysis was used to explore the mechanism of action of ZJZC in HLP treatment. The SwissTargetPrediction database was used to predict compound targets, and GeneCards, DisGeNet, OMIM, and DRUGBANK databases were used to identify HLP-related targets. Protein–protein interaction diagrams were constructed using the STRING database. The targets were subjected to gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The “herb-ingredient-target” network was visualized using Cytoscape. Preliminary validation was performed using molecular docking and enzyme-linked immunosorbent assay. Results: Ninety compounds were identified in ZJZC, including 34 flavonoids, 12 phenols, 10 terpenoids, 10 alkaloids, 8 organic acids, 8 anthraquinones, and 9 other compounds. In total, 904 targets were identified for these compounds. Among them, 158 targets intersected with the HLP target network. Network pharmacology analysis showed that MAPK1, PPAR-α, RXRA, HSP90AA1, PIK3R1, AKT1, PIK3CA, IL6, TNF, and ESR1 are the key targets of action. KEGG enrichment analysis identified 164 pathways. Among these, the AGE-RAGE signaling pathway in diabetic complications, lipid and atherosclerosis pathways, regulation of lipids in adipocytes, and insulin resistance are related to HLP. Molecular docking showed good affinity between the key targets and ingredients. Further, ZJZC treatment in mice resulted in lower expression of MAPK1 protein and increased expression of PPAR-α protein, which have been shown to be strongly associated with HLP. Conclusions This study showed that ZJZC contains various active ingredients and can modulate multiple targets and pathways associated with HLP, providing evidence at the molecular level for its clinical application in the treatment of HLP.

Recent innovations in nanomaterials inspire abundant novel tumor-targeting CRISPR-based gene therapies. However, the therapeutic efficiency of traditional targeted nanotherapeutic strategies is limited by that the biomarkers vary in a spatiotemporal-dependent manner with tumor progression. Here, we propose a self-amplifying logic-gated gene editing strategy for gene/H₂O₂-mediated/starvation multimodal cancer therapy. In this approach, a hypoxia-degradable covalent-organic framework (COF) is synthesized to coat a-ZIF-8 in which glucose oxidase (GOx) and CRISPR system are packaged. To intensify intracellular redox dyshomeostasis, DNAzymes which can cleave catalase mRNA are loaded as well. When the nanosystem gets into the tumor, the weakly acidic and hypoxic microenvironment degrades the ZIF-8@COF to activate GOx, which amplifies intracellular H⁺ and hypoxia, accelerating the nanocarrier degradation to guarantee available CRISPR plasmid and GOx release in target cells. These tandem reactions deplete glucose and oxygen, leading to logic-gated-triggered gene editing as well as synergistic gene/H₂O₂-mediated/starvation therapy. Overall, this approach highlights the biocomputing-based CRISPR delivery and underscores the great potential of precise cancer therapy.

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:Exploration of epidermal growth factor receptor (EGFR) expression and its clinical significance in pancreatic adenosquamous carcinoma (PASC).Methods:A total of 60 pancreatic cancer tissue samples and 8 normal pancreatic tissue samples were obtained from patients who were surgically treated at Beijing Chao-Yang Hospital, Capital Medical University from January 2016 to December 2021. A retrospective analysis of the clinical and pathological data of these 60 patients was conducted, including 23 males and 37 females with an age of (62.7±10.2) years. Among them, 20 cases were pathologically diagnosed as PASC, and 40 contemporaneous cases of pancreatic ductal adenocarcinoma (PDAC) were selected through propensity score matching. Immunohistochemistry (IHC) staining was used to measure the integrated optical density (IOD) of EGFR expression, and quantitative polymerase chain reaction (qPCR) was employed to detect the expression differences of EGFR mRNA. Based on the median IOD value of EGFR, the 20 PASC samples were divided into two groups, high and low expression groups. Kaplan-Meier survival analysis was performed to compare the impact of EGFR expression on the prognosis of PASC patients.Results:The IOD value of EGFR in PASC group (29.2 [25.7, 35.1]) was significantly higher than that in the PDAC group [9.5 (5.5, 13.0)] and they both exceeded the value in normal tissues [2.4 (1.7, 3.1)], with statistical significances ( all P<0.001 ). The level of EGFR mRNA expression in the PASC group was higher than that in the PDAC group [3.0 (1.8, 3.5) vs 1.2 (0.8, 1.2)], showing statistically significant difference ( P=0.0079). Patients with high EGFR expression had shorter overall survival compared with patients with low expression ( P=0.002). The incidence of vascular invasion in the PASC group [40.0% (8/20)] was higher than that in the PDAC group [17.5% (7/40)], with a significant difference ( P=0.002). The median survival time for the PASC group was 16.00 (9.25, 25.25) months, which was shorter than that of the PDAC group 21.50 (11.25, 40.75) months, showing a statistically significant difference ( P=0.033). The overall survival rate of the PASC group was lower than PDAC group ( P=0.028). Conclusion:EGFR expression is significantly elevated in PASC tissues and PASC patients have poor prognosis.

Objective:To investigate the feasibility of using anastomotic stoma as a long-term vascular access to maintain hemodialysis in patients who cannot establish an effective vascular access after ACUSEAL graft occlusion.Methods:A rare case of brachial artery-right atrium ACUSEAL artificial blood vessel fistula bypass to establish vascular access occlusion, the use of artificial blood vessel anastomosis stoma to establish buttonhole puncture as a long-term vascular access and the corresponding nursing methods.Results:At present, the patients were treated with regular dialysis for 32 months and blunt needle puncture for 23 months.Conclusions:Brachial artery-right atrium ACUSEAL graft is a rare vascular access surgery. As a new type of access with depleted autologous vascular resources, the efficacy is not certain. Because it is different from the traditional vascular prosthesis surgery, once the thrombotic occlusion of the graft fails, the risk and difficulty of reopening by interventional or surgical methods are great. Therefore, it is a challenging and reasonable method to use the limited residual cavity of the anastomosis as a long-term vascular access after occlusion to prolong the service life of the graft.

Objective:To investigate the emergency treatment and clinical effect when the guidewire stuck in the right atrium during central vein catheter placement for hemodialysis.Methods:Five cases with guidewire stuck in the right atrium during central vein catheter placement for hemodialysis from January 2011 to July 2018 admitted into the First Affiliated Hospital of Zhejiang University were retrospectively analyzed. In two cases, the guidewires were found completely stuck when the insert depth was about 20 cm. The guidewires were not able to move forward nor backward. In the other three cases, the guidewires could be moved forward but not backward with the insert depth at about 18 cm. All patients received emergent computed tomography angiography (CTA) or digital subtraction angiography (DSA) imaging. Images showed that the guidewires were stuck in the right atrium near the ventricular valve. The guidewire core drawing method, the multipurpose angiography(MPA) catheter capturing method and the manual guidewire adjusting method were used for emergent treatment.Results:One patient with completely stuck guidewire was successfully treated with guidewire core drawing method and the temporary central vein catheter catheterization through the internal jugular vein was performed under DSA. In a completely stuck case and a retrogradely stuck case, the J-shaped ends of the warped guidewires were captured into the MPA catheter, and the guidewires were then withdrawn from right atriums along with the contrast catheter. In the other 2 retrogradely stuck cases, under DSA, the guidewires were repeatedly pushed, the direction of J-shaped ends was manually adjusted, and then the guidewires were repeatedly pushed and pulled until catheters can be pulled out of the right atriums. The later 4 cases had permanent central vein catheter placement with the same guidewire after the stuck guidewires were withdrawn from the right atrium and readjusted.Conclusions:All three methods can successfully solve the emergent situation of the stuck guidewire in the right atrium. For patients with completely stuck guidewires, the MPA catheter capturing method can be simpler, safer, and more effective.

Objective To investigate the application of percutaneous transluminal balloon dilatation (PTA) in catheter replacement within patients with catheter-related central vein disease (CVD). Methods Thirteen cases of CVD patients from Jan 2015 to Mar 2018 admitted into the First Affiliated Hospital of Zhejiang University were retrospectively analyzed. All of them underwent digital subtraction angiography (DSA) to clarify problem origin. Suitable balloons were chosen to dilate the original catheters or the occlusive veins, and then the original catheters were replaced. PTA was used to help catheter replacement and all patients were followed up for 6 months. Results Four of the 13 patients were found stuck when replacing catheters. All of them successfully had catheters removed with PTA (Hong's techniques). All patients had successfully catheter replacement with blood flow volume>250 ml/min. Among 4 patients with edema, 3 patients showed better within 6 months. Only 3 patients needed warfarin to keep blood flow volume>250 ml/min within 6 months. Conclusions PTA shows advantages of lesser trauma, better tolerance and higher success rate in patients with catheter-related CVD. It can also relieve symptoms resulting from occlusive central vein.

We aimed to obtain the recombinant aminopeptidase encoded by Listeria monocytogenes (L. monocytogenes) gene lmo1711, and characterized the enzyme. First, the amino acid sequences of Lmo1711 from L. monocytogenes EGD-e and its homologues in other microbial species were aligned and the putative active sites were analyzed. The putative model of Lmo1711 was constructed through the SWISS-MODEL Workspace. Then, the plasmid pET30a-Lmo1711 was constructed and transformed into E. coli for expression of the recombinant Lmo1711. The his-tagged soluble protein was purified using the nickel-chelated affinity column chromatography. With the amino acid-p-nitroaniline as the substrate, Lmo1711 hydrolyzed the substrate to free p-nitroaniline monomers, whose absorbance measured at 405 nm reflected the aminopeptidase activity. The specificity of Lmo1711 to substrates was then examined by changing various substrates, and the effect of metal ions on the catalytic efficiency of this enzyme was further determined. Based on the bioinformatics data, Lmo1711 is a member of the M29 family aminopeptidases, containing a highly conserved catalytic motif (Glu-Glu-His-Tyr-His-Asp) with typical structure arrangements of the peptidase family. The recombinant Lmo1711 with a size of about 49.3 kDa exhibited aminopeptidase activity and had a selectivity to the substrates, with the highest degree of affinity for leucine-p-nitroaniline. Interestingly, the enzymatic activity of Lmo1711 can be activated by Cd²⁺, Zn²⁺, and is strongly stimulated by Co²⁺. We here, for the first time demonstrate that L. monocytogenes lmo1711 encodes a cobalt-activated aminopeptidase of M29 family.

Objective To investigate the clinical distribution and drug resistance of carbapenemsresistant Klebsiella pneumoniae (CRKP).Methods A total of 134 clinical strains of CRKP were collected from inpatients in our hospital from January 2014 to December 2016.VITEK-2 compact automatic microbiological analyzer was used to identify the bacteria and the supporting gram-negative bacterial drug susceptibility card was used for susceptibility testing.The sensitivity of other clinical commonly used antimicrobial agents was measured by K-B method.And the clinical distribution of CRKP and its resistance to antimicrobial agents were investigated and analyzed.Results The clinical strains of CRKP were isolated mainly from urine (n=70,52.2％) and sputum (n=38,28.4％).Klebsiella pneumoniae was identified in samples from the department of neurosurgery,including neurosurgery ICU (47.8％,64/134),ICU (23.9％,32/134),department of hepatobiliary surgery (8.2％,11/134) and department of urology (6.0％,8/134).Among all 22 antimicrobial agents tested,the resistant rates of CRKP to 16 antimicrobial agents were ＞90％,especially for ampicillin it was 100％,and those to 19 antibiotics were ＞80％,only for tigecycline it was 23.1％.There were 16 strains of CRKP with positive extended spectrum β-lactamases (ESBLs) (11.9％).All the CRKP strains were resistant to more than three kinds of antimicrobial agents,except one strain that was resistant to two kinds.Conclusion There is a wide range of clinical distribution of CRKP,which is resistant to most of antimicrobial agents,while tigecycline still has a strong antibacterial activity to CRKP.

Objective To investigate the effect of recombinant human interleukin 11(rhlL-11)in the treatment of idiopathic thrombocytopenic purpura(ITP)on the levels of Th1,Th2 and the expression of their transcription factors T-bet mRNA,GATA-3 mRNA.Methods Fifty-six cases adult ITP patients hospitalized in the department of hematology of the Second People's Hospital of Datong from May 2015 to December 2016 were collected,including 21 males and 35 females,aged 29~73 years; 10 healthy people in the same period were enrolled as control group,4 males and 6 females,aged 20~52 years.Th1 and Th2 cell ratio and Th1/Th2 ratio of ITP patients were detected by flow cytometry before and after treatment.The expression levels of transcription factor T-bet and GATA-3 were measured using real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-PCR)before and after treatment.Results The effective rate of rhlL-11 in ITP treatment was 76.8%(43/56).For the effective patients,the median PLT after treatment increased(25.0(15. 0,36.0)×109/L vs.68.0(49.0,108.0)×109/L,Z=-5.712,P<0.001); Th1 cells decreased,compared with that before the treatment(14.8 %(12.6%,17.6%)vs.10.6 %(9.8%,12.6%),Z=-4.825,P<0.001);Th2 cell increased,compared with that before the treatment(0.4%(0.3%,0.5%)vs.1.2%(0.9%,1.4%), Z=-5.720,P<0.001); Th1/Th2 decreased,compared with that before the treatment(40(30,49)vs.10.6(7.8,12.0),Z=-5.711,P<0.001];the expression level of T-bet mRNA decreased(0.36(0.18,0.51)vs 0.09(0.05,0.13),Z=-2.668,P=0.008);the expression level of GATA-3 mRNA increased,compared with that before treatment(0.04(0.03,0.05)vs.0.12(0.09,0.15),Z=-2.366,P=0.018).For ineffective patients,the median PLT before treatment was(11.0(8.0,15.5)×109/L),and the median PLT after treatment was(15.0(10.0,19.5)×109/L)(Z=-3.027,P=0.002); there was no significant difference in Th1,Th2, ratio of Th1/Th2 and T-bet and GATA-3 mRNA expression level before and after treatment in patients with ITP (P>0.05).Conclusion rhIL-11 can effectively correct the imbalance in Th1 and Th2 cells and the imbalance of T-bet and GATA-3 in ITP patients,but it has no obvious therapeutic effect on a small number of patients