Objective: To establish the quality standard for Yuhuang sprays. Methods: Berberine hydrochloride in Yuhuang sprays was qualitatively identified by TLC and determined by UV. The Vanillin-H2 SO4 method was employed for the determination of condensed tannins. Results:The spots obtained from the sample solutions showed the same color as those from the reference substance solution without interference from the negative sample. The calibration curve of berberiny hydrochloride was linear within the range of 1. 13-13. 60μg·ml-1(r=0. 999 9), and the average recovery was 99. 68% with RSD of 1. 53%(n=9). The calibration of catechin was linear within the range of 0. 03-0. 30 mg· ml-1(r=0. 999 3), and the average recovery was 99. 6% with RSD of 1. 76%(n=9). Conclusion:The method can be used in the quality control of Yuhuang sprays.

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 To evaluate the image quality improvement on abdominal CT imaging by using new model-based iterative reconstruction (MBIRn)in comparison with adaptive statistical iterative reconstruction (ASiR).Methods 40 patients who underwent upper abdominal three-phase contrast-enhanced scan were included.After scanning,all the scans obtained at 180 s later injection were reconstructed by three protocol,including ASiR (combined reconstruction of 40%FBP and 60%ASiR),the MBIRn of the noise reduction settings (MBIRNR40)and the spatinal resolution settings(MBIRSTND).The thickness of the slice was 0.625 mm.The values of CT and SD of the subcutaneous fat,left erector spinae,inferior vena cava and hepatic vein (left and right branches)were measured at the branch level of hepatic vein,and the contrast noise ratio (CNR)between inferior vena cava and hepatic vein were calculated.The subj ective image quality was evaluated by two radiologists according to the noise,smoothness and small branches of the inferior vena cava and hepatic vein using 5-scoring method.The quality images obtained from ASiR method were treated as reference standard.Results For MBIRSTNDand MBIRNR40images,the subjective noise decreased and image quality increased comparing with ASiR images.Among which the MBIRNR40images had the best image with vascular smoothness score and the lowest subjective noise.Conclusion Compared with ASiR,MBIRSTNDand MBIRNR40,especially MBIRNR40improves the quality of CT images of the inferior vena cava and its branches.

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 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.

Objective:To investigate the effects of poly(A) tails with different lengths on mRNA expression in vitro and the passage stability of transcription template with poly (A) tail in Escherichia coli ( E. coli). Methods:Plasmids with poly(A) tails of 38, 60, 103, 125 and 126 (60 nt+ 6 nt spacer+ 60 nt) nt were designed and constructed. Then the plasmids were linearized by single enzyme digestion and used as transcription template for preparing enhanced green fluorescent protein (EGFP)-mRNA. EGFP-mRNA containing poly(A) tails of different lengths were transfected into 293T cells and the expression of EGFP was detected by flow cytometry. As to stability test, the template plasmids with poly (A) tail of 125 and 126 nt were transformed into E. coli TransStbl3 and Top10 competent cells. Seven clones were selected for culture and plasmid extraction, and then the plasmids were digested by restriction enzyme and detected by capillary electrophoresis. For passage stability, three correctly sequenced clones of each group were selected for continuous passage at 37℃, and the plasmids were extracted and digested every two generations for capillary electrophoresis. At the same time, the correctly sequenced clones of 125 nt group were also passaged at 30℃, and the plasmids were also extracted and digested every two generations for capillary electrophoresis. Results:The transcription templates with poly(A) tail of different lengths were successfully constructed. Flow cytometry showed that the fluorescence expression of the template plasmids with poly (A) tail of 103 and 125 nt were significantly higher than that of 38 and 60 nt. The fluorescence expression of the plasmid with poly (A) tail of 126 nt was significantly higher than that of all other groups. The percentages of stable sequences of the template plasmid with poly(A) tail of 125 nt in TransStbl3 and Top10 competent cells were 76% and 91%, respectively. The results of continuous passage showed that poly(A) tail of 125 nt could be stable to the 4th generation at 37℃ in both TransStbl3 and Top10 competent cells, and stable to the 16th and 10th generations at 30℃. The percentages of stable sequences of the template plasmid with poly(A) tail of 126 nt in TransStbl3 and Top10 competent cells were 95% and 48%, respectively. The results of continuous passage showed that poly(A) tail of 126 nt could be stable to the 12th generation at 37℃ in both TransStbl3 and Top10 competent cells.Conclusions:The length and composition of poly(A) tail in mRNA affected the expression of target protein. Adding a spacer with a length of 6 nt to poly(A) tail and low temperature culture were both helpful to improve the stability of the template plasmid, which provided a reference for the design and preparation of in vitro transcription template of mRNA vaccine.

Blood stasis is an important pathological factor throughout the whole course of radiation-induced pulmonary fibrosis， which could evolve from new into long stagnation， and the methods of dispelling stasis to promote regeneration should throughout the whole disease progress. It is believed that the basis of the radiation-induced pulmonary fibrosis is heat toxin dispersing qi and yin， and deficiency of healthy qi promoting blood stasis. The process of the disease showed latent fire burning pulmonary collaterals， and the binding of phlegm and stasis. The key factors of the disease were the damage of ying-wei （营卫） qi in channels and collaterals， as well as the blood stasis evolving into dried blood. It is suggested that during radiotherapy， we should pay more attention to relieve heat， moisten dryness， supplement qi and yin， nourish and harmonize blood， and remove blood stasis， so as to prevent disease before it arises. If there is radiation pneumonia， we could focus on dissolving phlegm， removing blood stasis， clearing latent fire， and unblocking the collaterals and veins， in order to "control the development of existing disease". If it develops into radiation-induced pulmonary fibrosis， we could relive the center and supplement deficiency， tonify original qi， dispel stasis to promote regeneration， and clear dried blood， for the purpose of slowing the progression of disease. These ideas might provide reference for clinical treatment.

Objective To comprehensively review the clinical research on the treatment of hyperlipidemia with traditional Chinese Medicine(TCM)through the evidence mapping,and to understand the distribution of evidence in this field. Methods Databases including CNKI,Wangfang,VIP,SinoMed,PubMed,Cochrane Library,and Embase were searched from January 2004 to December 2023 to collect clinical studies,systematic reviews/meta-analyses,guidelines and clinical pathways related to the treatment of hyperlipidemia with TCM. The results were analyzed and displayed in charts and graphs according to the screening criteria,and the Assessment of Multiple Systematic Reviews 2 (AMSTAR 2) tool and the Preferred Reporting Item for Systematic Review and Meta-analysis of Chinese herbal medicine (PRISMA-CHM) were used to evaluate the quality of the systematic review/meta-analysis. Results A total of 1223 studies were included in the analysis according to Population,Intervention,Comparison,Outcome and Study design(PICOS) principles,involving 920 RCTs,249 non-RCTs,49 systematic reviews/meta-analyses,and 5 guidelines/expert consensus. In recent years,the overall number of clinical research publications has shown a downward trend. Hyperlipidemia frequently occurs in middle-aged and elderly people,and age of onset tends to be younger. The sample size of randomized controlled studies is mostly concentrated in 60-300 cases. There are many types of clinical treatment regimens for the treatment of hyperlipidemia with TCM,among which TCM decoction (50.13％) and Chinese patent medicine (38.41％) account for a relatively high proportion,and TCM exercise therapy (0.51％) is the lowest treatment. Jiangzhi Decoction has attracted more attention in trial group of TCM decoction,while Xuezhikang Capsule has attracted more attention in trial group of Chinese patent medicine. In terms of methodological design,199 papers(21.63％) explicitly mentioned the method of generating random sequence,17 papers(1.85％) mentioned allocation concealment,37 papers (4.02％) mentioned blinding. The control group was dominated by the statins,including simvastatin and atorvastatin. The outcome indicators mainly include the total effective rate,TCM syndrome score,blood lipid level,coagulation index,and adverse reactions,while the attention of TCM characteristic efficacy,inflammation,oxidative stress,and vascular endothelial index were low. The methodological and reporting quality of the systematic review/Meta-analysis were generally not high. AMSTAR-2 evaluation was extremely low,and the average PRISMA-CHM score was 15. Conclusion TCM has certain advantages in the treatment of hyperlipidemia,but there is a lack of high-quality evidence-based proof,and more high-quality clinical studies are still needed to further provide evidence supports in the future. It has been suggested that more large-sample and multi-center clinical studies should be carried out in the future. We should formulate systematic reviews/Meta analysis and guidelines/expert consensus according to the guidelines of clinical practice issues,also consult international standards and regulations,enhance normativity and reliability to improve the quality of their evidence.

ObjectiveThis study aims to investigate the effectiveness and potential mechanism of Erxian Decoction （二仙汤） for late-onset depression （LOD）. MethodsForty Wistar male rats of 7-8 weeks were divided into 20 each of normal group and youth depression group. Sixty Wistar male rats of 20 months were divided into 20 each of elder group， LOD group and Erxian Decoction group. The rats in youth depression group were modelled with chronic unpredictable mild stress （CUMS） for 6 weeks to build a depression model， and the elder rats in LOD group and Erxian Decoction group were modelled with CUMS for 6 weeks to build a LOD model， with no interventions in the normal group and the elder group. Gastric administration was carried out at the same time of modelling， rats in Erxian Decoction group were given 8 g/（kg·d） of Erxian Decoction by gavage， and rats in the normal group， youth depression group， elder group， and LOD group were given 4 ml/（kg·d） of pure water by gavage， for 6 weeks in each group. Sugar water preference experiment and forced swimming experiment were used to evaluate the depression-like behaviour of rats， absent field experiment was used to evaluate the spontaneous activity ability of rats， and T-maze experiment was used to evaluate the cognitive function of rats； Western blot assay was used to detect neuronal nuclei （NeuN）， nestin， doublecortin （DCX） in hippocampal tissue， and zonula occludens-l （ZO-1）， folate receptor α （FRα）， reduced folate carrier （RFC）， and protoncoupledfolate transporter （PCFT） protein expression in choroid plexus； immunofluorescence was used to detect the number of double-positive cells for Ki-67/nestin， the number of double-positive cells for 5-bromodeoxyuracil nucleoside （BrdU）/DCX in hippocampal dentate gyrus， and the protein expression of ZO-1， FRα， RFC， and PCFT in choroid plexus tissues； ELISA technique was used to detect plasma， cerebrospinal fluid， 5-methyltetrahydrofolate （5-MTHF） in hippocampal tissues； Pearson correlation analysis was used to correlate the 5-MTHF content in cerebrospinal fluid and hippocampal tissues and the expression of nestin， DCX， and NeuN proteins in hippocampal tissues of rats in youth depression group and LOD group. ResultsCompared with normal group， rats in youth depression group and LOD group showed reduced sugar water preference， reduced total distance travelled in the absent field， reduced number of times through the grid， prolonged forced swimming immobilisation， reduced hippocampal NeuN， nestin and DCX protein expression， reduced number of Ki-67/nestin double-positive cells in hippocampal dentate gyrus， reduced number of BrdU/DCX double-positive cells in hippocampal dentate gyrus， and reduced expression of ZO-1 and FRα proteins and fluorescence intensity （P＜0.05 or P＜0.01）； the correct rate of T-maze on days 3 and 4 in the rats of youth depression group and on days 2 to 4 in the rats of LOD group significantly decreased， and the content of 5-MTHF in the cerebrospinal fluid and hippocampal tissues of the rats of LOD group significantly decreased as well （P＜0.05 or P＜0.01）. Compared with youth depression group， rats in LOD group showed a decrease in total distance travelled in absenteeism， number of times through the grid， a significant decrease in the correct rate of the T-maze on day 1-4， a decrease in NeuN， nestin， DCX protein expression of hippocampal tissue and the number of Ki-67/nestin double-positive cells， BrdU/DCX double-positive cells of hippocampal dentate gyrus （P＜0.05 or P＜0.01）. Compared with LOD group， rats in Erxian Decoction group had elevated sugar-water preference and total distance travelled in absenteeism and number of times through the grid， shorter forced swimming immobility time， significantly higher correct rate of the T-maze on day 3-4， elevated expression of NeuN， nestin， and DCX proteins in hippocampal tissues， increased number of Ki-67/nestin double-positive cells and BrdU/DCX double-positive cells in hippocampal dentate gyrus， and increased 5-MTHF content in cerebrospinal fluid and hippocampal， and ZO-1， FRα， RFC， PCFT protein expression and fluorescence intensity increased in choroid plexus （P＜0.05 or P＜0.01）. Correlation analysis showed that there was a positive correlation between 5-MTHF content in cerebrospinal fluid （r = 0.466）， hippocampal tissue （r = 0.522） and nestin expression in hippocampal tissue （P＜0.05）. ConclusionErxian Decoction could improve depressive-like behaviour and cognitive impairment in LOD model rats， and its mechanism may promote hippocampal neurogenesis by alleviating structural damage to the choroid plexus of the brain tissue， and improving impaired choroid plexus folate transport.