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: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.

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.

ObjectiveTo establish and validate a clinical prediction model for 1-year major adverse cardiovascular events（MACEs）risk after percutaneous coronary intervention （PCI） in coronary heart disease （CHD） patients with blood stasis syndrome. MethodThe consecutive CHD patients diagnosed with blood stasis syndrome in the Department of Integrative Cardiology at China-Japan Friendship Hospital from September 1， 2019 to March 31， 2021 were selected for a retrospective study， and basic clinical features and relevant indicators were collected. Eligible patients were classified into a derivation set and a validation set at a ratio of 7∶3， and each set was further divided into a MACEs group and a non-MACEs group. The factors affecting the outcomes were screened out by least absolute shrinkage and selection operator （Lasso） and used to establish a logistic regression model and identify independent prediction variables. The goodness-of-fit of the model was evaluated by the Hosmer-Lemeshow test， and the area under curve （AUC） of the receiver operating characteristic （ROC） curve， calibration curve， decision curve analysis （DCA）， and clinical impact curve （CIC） were employed to evaluate the discrimination， calibration， and clinical impact of the model. ResultA total of 731 consecutive patients were assessed and 404 eligible patients were enrolled， including 283 patients in the derivation set and 121 patients in the validation set. Lasso identified ten variables influencing outcomes， which included age， sex， fasting plasma glucose （FPG）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， homocysteine （Hcy）， brachial-ankle pulse wave velocity （baPWV）， flow-mediated dilatation （FMD）， left ventricular ejection fraction （LVEF）， and Gensini score. The multivariate Logistic regression preliminarily identified age， FPG， TG， Hcy， LDL-C， LVEF， and Gensini score as the independent variables that influenced the outcomes. Of these variables， male， high FMD and high LVEF were protective factors， and the rest were risk factors. The prediction model for 1-year MACEs risk after PCI in CHD patients with blood stasis syndrome showed χ²=12.371 （P=0.14） in Hosmer-Lemeshow test and the AUC of 0.90. With the threshold probability > 10%， the model showed better prediction performance for 1-year MACEs risk after PCI in CHD patients with blood stasis syndrome than for that in all the patients. With the threshold probability > 60%， the estimated value was much closer to the real number of patients. ConclusionThe established clinical prediction model facilitates the early prediction of 1-year MACEs risk after PCI in CHD patients with blood stasis syndrome， which can provide ideas for the precise treatment of CHD patients after PCI and has guiding significance for improving the prognosis of the patients. Meanwhile， multi-center studies with larger sample sizes are expected to further validate， improve， and update the model.