BACKGROUND:It is confirmed that collagen sponge prepared from human tendon,bovine tendon,rat tail,pig skin and newborn bovine tendon have good cytocompatibility.OBJECTIVE:To extract collagen from newborn bovine skin,prepare the collagen sponge for biomedical application,and observe the biocompatibility and cytocompatibility of collagen sponge with lamb fibroblasts.DESIGN,TIME AND SETTING:Controlled study was performed in the Key Laboratory of Bioengineering of State Ethical Committee,Life Science and Engineering College,Northwest University for Nationalities from May 2006 to February 2007.MATERIALS:Newborn Galiba bovine within 24 hours and black fur lamb kidney fibroblasts were used.METHODS:Newborn bovine skin was harvested to prepare the collagen sponge with a series of procedures,including depilation,pepsin+glacial acetic acid,salting-out,dialysis and freeze drying.The obtained collagen sponge was inoculated with fibroblast suspension,which were divided into collagen sponge group,negative control group (saline) and positive control group (rubber bung leaching liquor).MAIN OUTCOME MEASURES:Inverted phase contrast microscope and JVC digital camera system were used to observe the cell morphology and growth.Acridine orange dyeing was used to observe the proliferation of cell in collagen sponge at 20 and 35 days of culture.Hematoxylin-eosin staining was used to observe the growth of cells in collagen sponge at 65 days of culture.RESULTS:The cells of positive control group were not adhesive and all died three days later.Those of collagen sponge group and negative control group were normal and adhesive.With the prolong of culture time,the sponge pore decreased gradually,sponge appearance became eminent and transparent,the cell increased in number but decreased in morphology.Acridine orange dyeing at 20 and 35 days of culture showed that a large amount of cells appeared in the co-culture of collagen sponge with lamb kidney fibroblast,and pack of cell clumps grew.Abundant blue nuclei and newborn red collagen fiber were found by hematoxylin-eosin staining.CONCLUSION:The collagen sponge from newborn bovine skin has a good biocompatibility with lamb kidney fibroblast cell of black fur,and no cytotoxicity appears.

BACKGROUND:Industrialization of new-born bovine serum and abundant resource of bovine lendon enable industrialization of medical collagen sponge.OBJECTIVE:To prepare collagen sponge with new-born bovine tendons by inoculating Veto cells,primary embryo skin cell of Tianzhu White Yak and lamb testicle cell of Minxian black fur sheep of the tissue scaffold of collagen sponge.and observe the biocompatibility of collagen sponge with three different cells.DESIGN,TIME AND SETTING:Repetitive measurement was performed at the Key Laboratory of State Nationalities Afrairs Committee,College of Life Sciences and Engineering,Northwest University for Nationalities from February to May 2006.MATERIALS:Tendons of new-born bovine within 24 hours were digested by glacial acetic acid and pepsinum firstly and then salting-out,dialysis and vacum freeze drying were performed to prepare collagen sponge.f2 passage of embryo skin cells of Tianzhu Whit Yak and f2 passage of lamb testicle cells of Minxian black fur sheep were prepared by out laboratory;Vero cells were provided by Union Medical University.METHODS:In a 6-hole plate,the Vero cell,embryo skin cells of Tianzhu White Yak and lamb testicle cells of Minxian Black Fur Sheep were inoculated into the collagen sponge pretreated by ultraviolet and sterilized by ozone.and incubated in 5%CO2 at 37℃.In addition,cells only inoculated ia a culture plate served as control. MAIN OUTCOME MEASURES:Inverted phase contrast microscope was used to observe cell growth condition in the collagen sponge and 6-hole plate at 5,12,24 and 72 hours.In addition,Coomassie brilliant blue as well as HE staining were conducted at 11 days after culture to identify the culture.RESULTS:Five hours after inoculation,cell adherence and expansion was observed at the bottom of culture plate.and some of the cells showed division.On the surface of collagen sponge.a round cell arrangement was observed.After inoculation of 48-72 hours,monolayer was found at the bottom of the plate.On the 11th day of culture.Coomassie brilliant blue and HE staining of three kinds of cells showed there were lager amount of cells well grew in the holes of collagen sponge,and the collagen sponge turned to be eminent,transparent and tenacious.CONCLUSION:The collagen sponge made from new-born bovine tendons exhibit good biocompatibility with three kinds of cells from different animals and tissues,and can be served as culture seaffoId of skin cells,tenal ceils.and testicle cells.

Real world clinical diagnosis and treatment activity is a complicated decision-making task. The effective clinical cases of traditional Chinese medicine (TCM) of highly experienced physicians play an important role in the routine diagnosis and treatment and the formulation of medical knowledge . Based on TCM electronic medical record data, this paper proposed a decision support prototype system on TCM clinical diagnosis and treatment based on TCM effective clinical cases and case-based reasoning (CBR) algorithm, which is used to assist inexperienced clinicians to make more reliable clinical decisions, and thereafter to improve the clinical curative effectiveness. The system integrates TCM clinical cases data set from a TCM clinical data warehouse, and retrieves the similar cases based on CBR method. In particular, according to the underlying personalized diagnosis and treatment for patients in TCM, this system implemented a flexible diagnosis and treatment modification mechanism based on correlation analysis among symptoms, diagnoses (syndrome or pattern in TCM) and medicine. Finally, through a demonstration of clinical application, we made an initial evaluation of the usefulness and practical effects of the system.

Objective This study aims to examine the potential mechanism of Shenfu Yixin Granules on heart failure(HF)based on network pharmacology,molecular docking,and experimental verification.Methods(1)The active components of herbs in Shenfu Yixin Granules were screened and retrieved through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).PubChem database and Swiss Target Prediction platform were used to predict targets.GeneCards and OMIM databases were used to screen HF-related targets.The intersection of active ingredient targets of Shenfu Yixin Granules and HF-related targets was performed by using Venny 2.1.0 platform to obtain common targets,which were the potential targets for anti-HF effect of Shenfu Yixin Granules.The potential targets were imported into the STRING database to construct a protein-protein interaction(PPI)network and screen the core targets of Shenfu Yixin Granules for the treatment of HF.GO functional and KEGG pathway enrichment analysis of potential targets were carried out by using DAVID database.AutoDock Vina software was used for molecular docking validation of key active ingredients and core targets.(2)SD rats were randomly allocated into sham operation group,model group,Shenfu Yixin Granules(5.28 g·kg-1)group,and positive control group(sacubitril-valsartan,20.8 mg·kg-1),with eight rats in each group.A rat model of HF after myocardial infarction was established by ligating the left anterior descending coronary artery.The rats were subsequently administered orally with the corresponding drugs once daily for a period of four weeks.Cardiac function including left ventricular ejection fraction(LVEF)and left ventricular fraction shortening(LVFS)in rats was assessed by echocardiography.Additionally,the histopathological alterations in rat heart tissue were examined using hematoxylin-eosin(HE)staining and Masson staining.The serum levels of brain natriuretic peptide(BNP),artial natriuretic peptide(ANP),and aldosterone(ALD)were determined by enzyme-linked immunosorbent assay(ELISA).Furthermore,real-time quantitative PCR and Western Blot were employed to detect mRNA and protein expressions of CAV1、F2 and MAPK1 in heart tissue.Results(1)A total of 210 active ingredients and 1 196 targets of Shenfu Yixin Granules,as well as 801 HF-related targets were obtained.Venny 2.1.0 platform was used to acquire 97 potential targets(common targets)of Shenfu Yixin Granules for the treatment of HF.Key active ingredients,such as quercetin,luteolin,arachidonic acid,kaempferol,and tanshinaldehyde were screened by"drugs-active ingredients-disease-targets"network analysis.The core targets including MAPK1、F2、CAV1、EDN1 and GJA1 were identified through PPI network analysis.The potential targets are mainly concentrated in multiple biological processes,namely,the positive regulation of gene expression,cardiac development,and the positive regulation of MAPK cascade,and involve multi key pathways including MAPK signaling pathway,HIF-1 signaling pathway and PI3K/Akt signaling pathway etc.Good binding activities were observed between MAPK1,CAV1,EDN1,F2 and quercetin,luteolin,kaempferol,tanshinaldehyde,as well as MAPK1,F2 and arachidonic acid.(2)Compared with sham operation group,LVEF and LVFS of rats significantly reduced(P＜0.01),heart mass index obviously increased(P＜0.05)in the model group.Myocardial tissue appears obvious pathological damage,and the degree of interstitial fibrosis was serious.The collagen volume fraction of the heart significantly increased(P＜0.01).The levels of serum BNP,ANP and ALD significantly increased(P＜0.01).The mRNA and protein expressions of CAV1、F2 and MAPK1 in heart tissue significantly increased(P＜0.05,P＜0.01).Compared with the model group,LVEF and LVFS of rats obviously increased(P＜0.01),but the decrease in heart mass index was not significant(P＞0.05)in Shenfu Yixin Granules group and positive control group.The pathological damage in myocardial tissues was significantly improved,the degree of interstitial fibrosis was significantly reduced.The collagen volume fraction of the heart significantly decreased(P＜0.01).The levels of serum BNP,ANP and ALD significantly decreased(P＜0.01).The mRNA and protein expressions of CAV1、F2 and MAPK1 in heart tissue significantly decreased(P＜0.05,P＜0.01).Conclusion Shenfu Yixin Granules may improve heart function and myocardial fibrosis in heart failure rats through the interaction between the active ingredients(quercetin,luteolin,arachidonic acid,kaempferol,and tanshinaldehyde)and targets(MAPK1,F2,CAV-1,and EDN1),so as to regulate MAPK signaling pathway and PI3K/Akt signaling pathway.

Objective:To analyze the clinical features, risk factors and prognosis of IPA combined with lung infection, aiming to further improve clinicians' understanding and diagnosis and treatment of it.Methods:Patients with IPA admitted to the Second Affiliated Hospital of Zhejiang University School of Medicine from January 2013 to October 2021 were retrospectively enrolled, and their clinical data was collected from the electronic medical record, including demographic information, clinical characteristics, biochemical indicators, auxiliary examination, microbial data and prognostic indicators. Patients were divided into two groups of IPA with pulmonary infection and IPA alone, and the clinical features, risk factors and prognosis of IPA patients with pulmonary infection were compared and analyzed in comparison with IPA patients alone.Results:A total of 156 IPA patients were finally recruited, with an average age of (67.12±12.89) years old and a main male proportion of 69.20%. Among them, there were 86 cases (55.13%) with IPA with pulmonary infection and 70 cases (44.87%) with IPA alone. Half of the IPA patients with pulmonary infection were mixed with one pathogen. The main pathogen of mixed infection was bacteria (82.72%), whereas acinetobacter baumannii accounted for the most common pathogen(25.93%, 42/162). Multivariate logistic regression analysis found that mechanical ventilation ( OR 4.89, 95% CI 2.23-10.70) and prior neutropenia ( OR 6.41, 95% CI 1.33-30.93) were independent risk factors for the occurrence of IPA with pulmonary infection. Compared with IPA alone, IPA patients with pulmonary infection were more likely to develop septic shock(69.80% vs. 32.90%, P <0.05), and have longer lengthes of hospital stay [16.00(8.00,36.50) vs.13.50 (7.00,20.50)] and ICU stay[11.50(6.00,31.25) vs.8.50(1.75,11.00)], and mechanical ventilation days [12.00(6.75, 25.25) vs.8.00(2.00,10.00)], as well as a higher 28-day mortality (55.80% vs.35.70%) and in-hospital mortality (64.00% vs. 35.70%). Conclusions:IPA patients with pulmonary infection accounts for more than half of IPA patients. The main respiratory etiology of IPA with pulmonary infection is acinetobacter baumannii. The independent risk factors of IPA patients with pulmonary infection are mechanical ventilation and neutropenia. The prognosis of IPA patients with pulmonary infection is worse than patients with IPA alone, which is worthy for the attention of physicians.

Objective:To investigate the risk factors of proteinuria in patients with hypertension in Qinghai-Tibet Plateau.Methods:From March 2019 to June 2020, prospective design was used to collect data of Qinghai-Tibet Plateau hypertension patients who were eligible for continuous enrollment in the Department of Cardiovascular Medicine in Hospital of Chengdu Office of People's Government of Tibet Autonomous Region. Questionnaire survey, physical examination and blood pressure measurement were performed on the selected patients. Fasting venous blood samples were collected for liver function test, blood lipid test, blood glucose test, and hemoglobin test, etc. Three times of morning urine samples were taken on different days, and urine protein creatinine ratio (UACR) was measured, UACR < 30 mg/g was negative for urinary protein, and UACR≥30 mg/g was positive for urinary protein. At the same time, the selected patients were examined by carotid artery color ultrasound and heart color ultrasound. The risk factors of proteinuria were analyzed.Results:A total of 588 patients with hypertension met the inclusion criteria, including 472 patients (80.3%) who received antihypertensive drug therapy, 239 patients (40.6%) had antihypertensive treatment compliance, and 252 patients (42.9%) reached the standard blood pressure after theropy. Hypertension was associated with diabetes mellitus in 150 patients (25.5%), and urinary protein was positive in 126 patients (21.4%). In univariate analysis, ethnic composition, systolic blood pressure [(138.19 ± 19.65) vs (133.16 ± 18.45) mmHg, 1 mmHg = 0.133 kPa], diastolic blood pressure [(85.80 ± 13.51) vs (83.17 ± 12.19) mmHg], uric acid [(411.79 ± 101.54) vs (379.96 ± 102.18) μmol/L], hemoglobin [(152.86 ± 30.70) vs (143.49 ± 21.15) g/L], pulmonary artery trunk width [(21.76 ± 3.94) vs (20.98 ± 3.34) mm], and ventricular septal thickness [(9.90 ± 1.70) vs (9.47 ± 1.60) mm] in the positive group ( n = 126) were significantly higher than those in the negative group ( n = 462, P < 0.01 or < 0.05). In multivariate logistic regression analysis, increased systolic blood pressure [odds ratio ( OR) = 1.015, 95% confidence interval (95% CI): 1.005 - 1.026], uric acid ( OR = 1.003, 95% CI: 1.001 - 1.005), and pulmonary artery trunk width ( OR = 1.058, 95% CI: 1.001 - 1.118) were risk factors for proteinuria; Tibetans had a decreased risk of proteinuria compared with Han ( OR = 0.505, 95% CI: 0.317 - 0.805), but increased hemoglobin had an increased risk of proteinuria compared with normal hemoglobin ( OR = 1.890, 95% CI: 1.231 - 2.903). Conclusion:In patients with hypertension at high altitude, increased hemoglobin, systolic blood pressure, uric acid, pulmonary artery trunk width, and Han nationality are risk factors for proteinuria.

Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)-induced cytokine storms constitute the primary cause of coronavirus disease 19(COVID-19)progression,severity,criticality,and death.Gluco-corticoid and anti-cytokine therapies are frequently administered to treat COVID-19,but have limited clinical efficacy in severe and critical cases.Nevertheless,the weaknesses of these treatment modalities have prompted the development of anti-inflammatory therapy against this infection.We found that the broad-spectrum anti-inflammatory agent inosine downregulated proinflammatory interleukin(IL)-6,upregulated anti-inflammatory IL-10,and ameliorated acute inflammatory lung injury caused by mul-tiple infectious agents.Inosine significantly improved survival in mice infected with SARS-CoV-2.It indirectly impeded TANK-binding kinase 1(TBK1)phosphorylation by binding stimulator of interferon genes(STING)and glycogen synthase kinase-3β(GSK3β),inhibited the activation and nuclear trans-location of the downstream transcription factors interferon regulatory factor(IRF3)and nuclear factor kappa B(NF-κB),and downregulated IL-6 in the sera and lung tissues of mice infected with lipopoly-saccharide(LPS),H1N1,or SARS-CoV-2.Thus,inosine administration is feasible for clinical anti-inflammatory therapy against severe and critical COVID-19.Moreover,targeting TBK1 is a promising strategy for inhibiting cytokine storms and mitigating acute inflammatory lung injury induced by SARS-CoV-2 and other infectious agents.

Objective:To evaluate the immunogenicity of a quadrivalent subunit vaccine combined with RFH01 adjuvant in a mouse model.Methods:Identification tests were performed on four monovalent influenza virus subunit vaccine stock solutions according to the methods described in Part 3 of the Chinese Pharmacopoeia 2020 Edition. In the study of the quadrivalent subunit vaccine combined with RFH01 adjuvant, 460 female BALB/c mice (6-8 weeks old) were randomly divided into 46 groups including experimental groups, vaccine control group, negative control group and blank group with 10 mice in each group. In the study of the quadrivalent subunit vaccine in old and young mice, 80 female 10-month-old and 80 female 10-week-old BALB/c mice were randomly divided into 16 groups ( n=10) including monovalent influenza virus vaccine group, quadrivalent subunit vaccine group, quadrivalent subunit vaccine+ RFH01 adjuvant group, chicken embryo quadrivalent split vaccine control group and PBS group. All mice were immunized by intramuscular injection. At 21 d after the primary immunization, a booster immunization was conducted using the same strategy. Blood samples were collected at 21 d and 42 d after the primary immunization for serum separation. Haemagglutination inhibition (HI) test was performed to detect the antibody levels in mouse serum samples. Results:After the booster immunization, the positive conversion rates in all vaccine+ RFH01 adjuvant groups reached 100%, and the geometric mean titers (GMTs) of serum antibodies were significantly higher than those of the vaccine groups without RFH01 adjuvant. There were significant differences in serum antibody titers between the monovalent/quadrivalent subunit vaccine groups with and without RFH01 adjuvant. After the booster immunization, the titers of serum antibodies against H1N1, H3N2, B/Victoria and B/Yamagata in the 10-week-old mice were significantly higher than those in the 10-month-old mice.Conclusions:The monovalent and quadrivalent influenza virus vaccines in combination with RFH01 adjuvant could elicit higher antibody titers in young (6-10 weeks old) and old (10 months old) mice, showing good immunogenicity.

Human metapneumovirus(hMPV)is a common cause of acute lower respiratory infection(ALRI)in children, especially in children aged 2~5 years.The study and mastery of the epidemiological characteristics and transformation patterns of various subtypes of hMPV can lead to a deeper understanding of the distribution areas, epidemiological years and clinical relevance of various subtypes of hMPV.It is important to carry out systematic and comprehensive genotyping and epidemiological study of hMPV to reveal the distribution characteristics and epidemic trend of hMPV.In this review, the research progress in the epidemiology of hMPV is reviewed, which provides ideas for the surveillance, prevention and clinical treatment of hMPV infection, and provides reference for the development of hMPV vaccine and disease prevention and control.

Objective To investigate the effect of Ba Bao Dan(BBD)on cardiac injury induced by doxorubicin in zebrafish.Methods We induced a zebrafish myocardial injury model using the chemotherapeutic drug,doxorubicin.We then examined the effects of different concentrations of BBD on pericardial edema and heart rate under an in vivo microscope.We also examined the inhibitory effects of BBD on neutrophil infiltration in the heart in Tg(mpx:EGFP)transgenic zebrafish.The impacts of BBD on superoxide dismutase,catalase,and malondialdehyde were observed.mRNA expression levels of ferroptosis-related factors,including glutathione peroxidase 4a(gpx4a),prostaglandin-endoperoxide synthase 2(ptgs2),arachidonate 5-lipoxygenase(alox5a),and acyl-CoA synthetase long-chain family member 4(acsl4)were determined by Real-time quantitative polymerase chain reaction.The accumulation of ferrous ions in zebrafish heart was assessed using a fluorescent probe for ferrous ions.Results BBD alleviated doxorubicin-induced pericardial edema and bradycardia in zebrafish,reduced neutrophil infiltration in the heart(P＜0.05),decreased malondialdehyde concentration(P＜0.05),and enhanced the activities of superoxide dismutase and catalase(P＜0.05).BBD also significantly inhibited ferroptosis,reduced the accumulation of ferrous ions in the zebrafish heart,suppressed the expression of ptgs2,alox5a,and acsl4(P＜0.05),and promoted the expression of gpx4a(P＜0.05).Conclusions BBD can attenuate doxorubicin-induced zebrafish myocardial injury and improve cardiac function by inhibiting lipid peroxidation and regulating ferroptosis.