Objective To investigate the effects of neurotrophic factor Neuritin overexpression on the angiogenic effects of chicken embryonic allantoic membrane (CAM) and human umbilical vein endothelial cells (HUVECs),and to provide a new direction for the treatment of angiogenic diseases. Methods Thirty fresh yellow-skinned breeding eggs were selected to establish a CAM model,which were divided into three groups by randomized numerical table method:positive control group (bFGF),negative control group (NS) and experimental group (Neuritin),with 10 eggs in each group. The positive control group was loaded with 2500 U/mL of bFGF,the experimental group was loaded with 10 μg/mL of Neuritin protein,and the negative control group was loaded with NS. 10 μL loading volume was loaded into each group,and all CAMs were incubated at the same temperature,relative humidity,and time,and the vascular branching,number,and size of the CAMs in each group were recorded after 72 h of incubation. Fresh umbilical cords from healthy pregnant women were selected to produce primary HUVECs,which were divided into three groups:transfected with recombinant plasmid (HUVEC-neu group),transfected with empty vector (HUVEC-3.1 group),and untransfected (HUVEC group). Primary HUVECs in the HUVEC-neu group were transfected with the recombinant plasmid Neuritin,and those in the HUVEC-3.1 group were transfected with the empty vector. HUVEC-3.1 group was transfected with the empty vector plasmid,and HUVEC group was not given any special treatment,and all three groups received the same culture regimen. Western blot was used to detect the protein expression level of Neuritin in HUVEC-3.1 and HUVEC-neu groups. CCK-8 assay,cell scratch assay,Transwell assay,and tube formation assay were used to detect protein expression level of Neuritin in HUVEC-3.1 group and HUVEC-neu group,and HUVEC-neu groups for cell proliferation,migration and tube formation. Results (1) The number of CAM vessel branch points and microvessels in the experimental group was significantly increased compared with that in the negative control group (P<0.01),but there was no statistically significant difference in the number of large and medium-sized vessels between the two groups (P>0.05);(2) Neuritin was successfully overexpressed in HUVECs in the HUVEC-neu group. (3) Compared with the HUVEC-3.1 group,the proliferation vigor of cells in the HUVEC-neu group was decreased (P<0.05),but their migration and tube formation abilities were significantly enhanced (P<0.01). Conclusion Neuritin overexpression promotes angiogenesis and participates in the regulation of neovascularization by affecting cell prolif-eration,migration,and tube formation ability.

Objective: To investigate the influencing factors for delay in healthcare-seeking, definitive diagnosis and identification in patients with pulmonary tuberculosis (PTB) in Minhang District, Shanghai Municipality, so as to provide the basis for effectively reducing delay in PTB patients. Methods: Data of PTB patients in Minhang District from 2017 to 2022 were collected from the Infectious Disease Reporting Information System of Chinese Disease Prevention and Control Information System. The prevalence rates of delay in healthcare-seeking, definitive diagnosis and identification were analyzed, and factors affecting delay in healthcare-seeking, definitive diagnosis and identification were identified using multivariable logistic regression models. Results: A total of 4 214 PTB patients were reported in Minhang District from 2017 to 2022, including 2 802 males and 1 412 females, with a male-to-female ratio of 1.98∶1. The majority of patients were aged 25 to <45 years (1 664 cases, 39.49%). The prevalence rates of delay in healthcare-seeking, definitive diagnosis and identification were 36.81%, 30.21% and 38.09%, respectively. Delay in healthcare-seeking was associated with the year (2018, OR=0.708; 2019, OR=0.549; 2020, OR=0.670; 2021, OR=0.682), gender (female, OR=1.199), occupation (worker, OR=1.379; housekeeping service/housework/unemployed, OR=1.481), case identification route (symptom-based consultation, OR=11.159), and level of the first-diagnosed hospital (city-level, OR=1.528). Delay in definitive diagnosis was associated with age (45 to <65 years, OR=1.476), occupation (commercial service, OR=0.687; housekeeping service/housework/unemployed, OR=0.672), household registration (non-local, OR=0.820), case identification route (symptom-based consultation, OR=0.616), pathogen test result (negative/not tested, OR=1.903), and the level of the first-diagnosed hospital (city-level, OR=0.311). Delay in identification was associated with the year (2018, OR=0.785; 2019, OR=0.647; 2020, OR=0.790; 2021, OR=0.710), occupation (commercial service, OR=0.687), household registration (non-local, OR=0.848) and level of the first-diagnosed hospital (city-level, OR=0.560) Conclusions Year, gender, occupation, case identification route and level of the first-diagnosed hospital are influencing factors for delay in healthcare-seeking in PTB patients. Age, occupation, household registration, case identification route, pathogen test result and level of the first-diagnosed hospital are influencing factors for delay in definitive diagnosis. Year, occupation, household registration and level of the first-diagnosed hospital are influencing factors for delay in identification.

OBJECTIVE To study the effect and safety evaluation of percutaneous tracheotomy and traditional open tracheotomy in patients with severe pulmonary infection in ICU environment.METHODS A retrospective analysis was conducted on 80 patients with severe pulmonary infections admitted to the ICU of Shangqiu First People's Hospital from July 2019 to June 2022 as the subjects of the study.According to the tracheotomy approach,they were divided into the Percutaneous tracheotomy group(n=40)and the Conventional tracheotomy group(n=40).The Conventional tracheotomy group underwent traditional routine tracheotomy,while the Percutaneous tracheotomy group underwent percutaneous tracheotomy;all tracheotomies were performed by the same physician.The two groups were compared based on different tracheotomy techniques,and the surgical incision time,intraoperative blood loss,complete healing indicators were analyzed to evaluate the surgical outcomes and complication rates in each group.RESULTS Compared with the Conventional tracheotomy group,the Percutaneous tracheotomy group significantly reduced surgical time,intraoperative blood loss,and wound healing time after extubating(P<0.05),and the incidence of complications such as difficult cannulation,pneumothorax and subcutaneous emphysema decreased significantly,and the difference was statistically significant(P<0.05).In the BMI subgroup analysis of complications,it was found that in the BMI≥28.0 kg/m2 subgroup,the incidence of casing difficulty and pneumothorax complications in the Percutaneous tracheotomy group was lower than that in the Conventional tracheotomy group,and the difference was statistically significant(P<0.05).In the 22.0 kg/m2≤BMI<28.0 kg/m2 subgroup and BMI<22.0 kg/m2 subgroup,there was no significant difference in the difficulty of cannulation and pneumothorax complications between the two groups(P>0.05).CONCLUSION In ICU patients with severe pneumonia,early percutaneous tracheotomy can significantly reduce intraoperative bleeding blood volume,shorten the time of tracheotomy,especially in patients with BMI≥28.0 kg/m2,which has a lower incidence of complications.Percutaneous tracheotomy is worth promoting in ICU patients with severe lung infection who need tracheotomy.

Objective To investigate the effects of neurotrophic factor Neuritin overexpression on the angiogenic effects of chicken embryonic allantoic membrane (CAM) and human umbilical vein endothelial cells (HUVECs),and to provide a new direction for the treatment of angiogenic diseases. Methods Thirty fresh yellow-skinned breeding eggs were selected to establish a CAM model,which were divided into three groups by randomized numerical table method:positive control group (bFGF),negative control group (NS) and experimental group (Neuritin),with 10 eggs in each group. The positive control group was loaded with 2500 U/mL of bFGF,the experimental group was loaded with 10 μg/mL of Neuritin protein,and the negative control group was loaded with NS. 10 μL loading volume was loaded into each group,and all CAMs were incubated at the same temperature,relative humidity,and time,and the vascular branching,number,and size of the CAMs in each group were recorded after 72 h of incubation. Fresh umbilical cords from healthy pregnant women were selected to produce primary HUVECs,which were divided into three groups:transfected with recombinant plasmid (HUVEC-neu group),transfected with empty vector (HUVEC-3.1 group),and untransfected (HUVEC group). Primary HUVECs in the HUVEC-neu group were transfected with the recombinant plasmid Neuritin,and those in the HUVEC-3.1 group were transfected with the empty vector. HUVEC-3.1 group was transfected with the empty vector plasmid,and HUVEC group was not given any special treatment,and all three groups received the same culture regimen. Western blot was used to detect the protein expression level of Neuritin in HUVEC-3.1 and HUVEC-neu groups. CCK-8 assay,cell scratch assay,Transwell assay,and tube formation assay were used to detect protein expression level of Neuritin in HUVEC-3.1 group and HUVEC-neu group,and HUVEC-neu groups for cell proliferation,migration and tube formation. Results (1) The number of CAM vessel branch points and microvessels in the experimental group was significantly increased compared with that in the negative control group (P<0.01),but there was no statistically significant difference in the number of large and medium-sized vessels between the two groups (P>0.05);(2) Neuritin was successfully overexpressed in HUVECs in the HUVEC-neu group. (3) Compared with the HUVEC-3.1 group,the proliferation vigor of cells in the HUVEC-neu group was decreased (P<0.05),but their migration and tube formation abilities were significantly enhanced (P<0.01). Conclusion Neuritin overexpression promotes angiogenesis and participates in the regulation of neovascularization by affecting cell prolif-eration,migration,and tube formation ability.

Celastrol is extracted from the traditional Chinese medicine Tripterygium wilfordii Hook.f..It is a kind of traditional Chinese medicine monomer with extensive pharmacological activity and has anti-tumor,anti-inflammation,anti-oxidation and neuroprotective effects.Studies have found that celastrol is not only closely related to obesity,tumor and cardiovascular diseases,but also plays a neuroprotective role in the cerebrovascular system by regulating various signaling pathways.At present,effective drugs for stroke are still limited,but with the deepening of the research on celastrol,its therapeutic potential in stroke has received more and more attention,especially in ischemic and hemorrhagic stroke,which has shown good therapeutic effects.Therefore,this is the first time to systematically summarize the therapeutic effects of celastrol on stroke and the underlying mechanisms involved,in order to provide further directions and references for the neuroprotective effects of celastrol.

Background::There is a need for effective and safe therapies for psoriasis that provide sustained benefits. The aim of this study was to assess the efficacy and safety of tildrakizumab, an anti-interleukin-23p19 monoclonal antibody, for treating moderate-to-severe plaque psoriasis in Chinese patients.Methods::In this multi-center, double-blind, phase III trial, patients with moderate-to-severe plaque psoriasis were enrolled and randomly assigned (1:1) to receive subcutaneous tildrakizumab 100 mg or placebo at weeks 0 and 4. Patients initially assigned to placebo were switched to receive tildrakizumab at weeks 12, 16, and every 12 weeks thereafter. Patients in the tildrakizumab group continued with tildrakizumab at week 16, and every 12 weeks until week 52. The primary endpoint was the Psoriasis Area and Severity Index (PASI 75) response rate at week 12.Results::At week 12, tildrakizumab demonstrated significantly higher PASI 75 response rates (66.4% [73/110] vs. 12.7% [14/110]; difference, 51.4% [95% confidence interval (CI), 40.72, 62.13]; P <0.001) and Physician’s Global Assessment (60.9% [67/110] vs. 10.0% [11/110]; difference, 49.1% [95% CI, 38.64, 59.62]; P <0.001) compared to placebo. PASI 75 response continued to improve over time in both tildrakizumab and placebo-switching to tildrakizumab groups, reaching maximal efficacy after 28 weeks (86.8% [92/106] vs. 82.4% [89/108]) and maintained up to 52 weeks (91.3% [95/104] vs. 87.4% [90/103]). Most treatment-emergent adverse events were mild and not related to tildrakizumab. Conclusion::Tildrakizumab demonstrated durable efficacy through week 52 and was well tolerated in Chinese patients with moderate-to-severe plaque psoriasis.Trial registration::ClinicalTrials.gov, NCT05108766.

Objective The aim of this study was to evaluate the efficacy and side effects of sacubitril/valsartan in the treatment of patients with chronic kidney disease(CKD)at stage 5 with resistant hypertension,and to explore the cardiovascular benefits and security of medical in the patients.Methods Patients with CKD5 resistant hypertension diagnosed and treated in the First Affiliated Hospital of Guangxi Medical University from September 2020 to March 2022 were selected and divided into the observation group(treated with routine treatment of kidney disease at end-stage and sacubitril/valsartan)and control group(include droutine treatment of renal disease at end-stage and ACEI or ARB drugs)according to treatment strategy.The patients in both two groups were treated with adequate dialysis treatment and conventional drug treatment of renal disease at end-stage.The patients were followed up for at least 3 months,the clinical efficacy of three months after treated with sacubitril/valsartan was observed,and the efficacy indicators and security indicators and adverse cardiovascular events were observed,the occurrence of adverse effects during the period of drug use were compared with the control group.Results A total of 110 patients were included in this study and there were 55 cases in each group.There were no significant differences in gender,age,age of dialysis,etiology,dialysis mode and blood pressure between the two groups(P>0.05).The Systolic blood pressure(SBP),diastolic blood pressure(DBP),b-type urinary natriuretic peptide precursor(Pro-BNP)and cardiac function grade in the observation group after treatment was significantly decreased compared with before treatment.The left ventricular ejection fraction(LVEF)and the ratio of LVEF<50%in the observation group was significantly reduced after treatment(P<0.05).SBP,DBP and Pro-BNP decreased 3 months after treatment compared with the baseline before treatment,and improved significantly in the first month after treatment(P<0.05).The decrease of DBP and BNP before and after treatment was significantly different between the two groups,and the decrease of DBP and BNP was more significant in the observation group(P<0.05).The difference of LVEF and left ventricular end diastolic diameter(LVEDD)between the two groups before and after treatment was statistically significant,and the improvement was more obvious in the observation group(P<0.05).There were no significant differences in the safety indicators of serum potassium,estimated glomerular filtration rate(eGFR)and liver function between two groups before and after treatment(P>0.05).In terms of adverse reactions,only 1 case in the control group developed hyperkalemia within 3 months of follow-up,and no hypotension or other adverse reactions occurred in the two groups.Conclusions The treatment of patients with CKD stage 5 hypertension with sacubitril/valsartan has obvious cardiovascular benefits.Sacubitril/Valsartan has efficacy in lowering blood pressure,improving cardiac function and reducing volume load,with less adverse events and higher safety than control group.

Objective By assessing the uncertainty of the determination results for the concentration of ambroxol hydrochloride injection,the reliability of the concentration measurement results is improved,ensuring the safety and efficacy of high-risk dosage form medications.Methods The uncertainty assessment of the content determination results by HPLC external standard method was conducted using both the TOP-DOWN method and the GUM method in the past.The TOP-DOWN method evaluated after synthesizing two components:the combined bias uncertainty component and the intermediate precision uncertainty component.The GUM method analyzed six sources of uncertainty based on the experimental process and evaluated after synthesizing the uncertainty.Results When the confidence level was 95％,the expanded uncertainty assessed by the TOP-DOWN method and the GUM method was 1.84％and 2.36％respectively(K=2).Conclusion The results from both assessments indicated that the determination of ambroxol hydrochloride injection content was reliable,ensuring the safety and efficacy of the drug.

【Objective】 To establish an enzyme-linked immunosorbent assay (ELISA) method for the determination of residual human coagulation factor Ⅺ in human prothrombin complex and validate the method. 【Methods】 Human factor Ⅺ was reacted with the capture antibody coated on the microtiter plate. After appropriate washing steps, biotinylated primary antibody was bound to the captured protein. Excess primary antibody was washed away and bound antibody was reacted with horseradish peroxidase conjugated streptavidin. TMB substrate was used for color development at 450 nm. The dilution reliability, accuracy, specificity, repeatability, intermediate precision, linearity, range and durability were verified. 【Results】 The verification results showed that the accuracy and specificity of this method met the experimental requirements, with an average recovery rate of 109.2% and RSD of 6.93%. The repeatability RSD was 6.78%, and the intermediate precision RSD was 6.75%, indicating good precision. The linear regression correlation coefficient of standard curve was 0.999 9, showing good accuracy and precision within the linear range. The durability was verified by the incubation time and the validity period of reagent kit opening. The results showed that the RSD of the incubation time change was 6.62%, indicating that the incubation time of this detection method was controlled between 28 to 32 minutes, and there was no significant impact on the results. The RSD of the detection results before and after the reagent kit was opened and stored under conditions for 7 days was 3.84%, indicating that the preservation of the reagent kit according to the conditions for 7 days after opening has no effect on the FⅪ detection results. Both indicated that the method had good durability. The dilution reliability results showed that there was a "hook" effect in the detection of FⅪ residue in human prothrombin complex, which could be solved by diluting 100 to 200 times. 【Conclusion】 This method can be used for the determination of FⅪ residues of human prothrombin complex in laboratory.

Purpose: Gastric cancer (GC) has high morbidity and mortality, the cure rate of surgical treatment and drug chemotherapy is not ideal. Therefore, development of new treatment strategies is necessary. We aimed to identify the mechanism underlying Sp1 regulation of GC progression. Methods: and Methods: The levels of Sp1, β-catenin, SET domain bifurcated 1 (SETDB1), and 15-hydroxyprostaglandin dehydrogenase (HPGD) were detected by quantitative reverse transcription polymerase chain reaction and western blot analysis. The targets of SETDB1 were predicted by AnimalTFDB, and dual-luciferase reporter assay was used for confirming the combination of Sp1, β-catenin, and SETDB1. HGC27 or AGS cells (1×10 6 cells/mouse) were injected into mice via the caudal vein for GC model establishment. The level of Ki67 was detected using immunohistochemistry, and hematoxylin and eosin staining was performed for evaluating tumor metastasis in mice with GC. Results: HPGD was inhibited, while the protein levels of Sp1, β-catenin, and SETDB1 were up-regulated in GC tissues and cell lines. HPGD overexpression or SETDB1 silencing inhibited the proliferation, invasion, and migration of GC cells, and Sp1 regulated the proliferation, invasion, and migration of GC cells in a β-catenin-dependent manner. Furthermore, HPGD served as a target of SETDB1, and it was negatively regulated by SETDB1; additionally, Sp1 and β-catenin bound to the SETDB1 promoter and negatively regulated HPGD expression. We proved that Sp1 regulated GC progression via the SETDB1/HPGD axis. Conclusions Our findings revealed that Sp1 transcriptionally inhibited HPGD via SETDB1 in a β-catenin-dependent manner and promoted the proliferation and metastasis of GC cells.