Objective To study the correlation of an ultra high performance liquid chromatography(UPLC)fingerprint of Xiaochengqitang pieces(decoction and granules).Methods The UPLC method was used to establish the fingerprint of 15 batches of Xiaochengqitang pieces(decoction and granules).The correlation of the three UPLC fingerprints was evaluated by similarity analysis,pearson correlation analysis,cluster analysis(CA),principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA).Results UPLC fingerprints of 15 batches of Xiaochengqitang pieces(decoction and granules)determined 16 common peaks,and 14 peaks were identified.The similarity of the fingerprints of the 15 batches of Xiaochengqitang pieces(decoction and granules)with the corresponding control fingerprints was greater than 0.90,and the similarity of the three control fingerprints was greater than 0.88.The results of pearson correlation analysis showed that 8 common peaks in Xiaochengqitang pieces(decoction and granules)had a very significant positive correlation.The results of CA showed that the properties of Xiaochengqitang decoction and granules were more similar.The results of PCA showed that the principal components with 4 eigenvalues greater than 1 contained 88％of the information of the original data.OPLS-DA screened 7 differential markers with variable importance projection value greater than 1.Conclusion The main chemical compositions of Xiaochengqitang pieces(decoction and granules)are consistent,which can provide data support for the quality control and clinical use of Xiaochengqitang compound preparation.

Objective To establish a quantitative analysis of multi-compounds by single-marker(QAMS)method for the simultaneous determination of four compounds in formula granules of Pteris multifida and optimize the extraction process of total flavonoids.Methods The relative correction factors(RCFs)of lonicerin,luteolin and apigenin in the formula granules of Pteris multifida were calculated by using the UPLC method with rhoifolin as the internal reference,and their durability was investigated.The external standard method(ESM)was used to determine the content of four compounds in the formula granules of Pteris multifida,and the difference between the calculated values and the measured values was compared.The effects of ultrasonic time,ethanol volume fraction,solid-liquid ratio and ultrasonic power on the extraction rate of total flavonoids were studied by single factor experiment.On this basis,Box-Behnken test with three factors and three levels was conducted to optimize the extraction process of total flavonoids,and the verification experiment was conducted.Results The content of rhoifolin in the formula granules was 0.035%-0.056%,and the content of lonicerin,luteolin and apigenin by QAMS method was 0.085%-0.167%,0.014%-0.028%and 0.004%-0.008%,respectively,which had no significant difference with the external standard method.The optimal extraction conditions of total flavonoids were 80%ethanol water,solid-liquid ratio 1:20,40 minutes extraction,the average extraction rate was 24.46 mg·g-1.Conclusion The established QAMS method was accurate and feasible,and the optimized extraction process of total flavonoids based on Box-Behnken response surface method was simple and feasible,which could lay a foundation for the quality evaluation of the Pteris multifida formula granules.

Objective: To explore the current status related to behavioral problems among children and adolescents aged 12-16 years old in Guizhou Province, China, and to provide a reference for timely prevention and intervention measures. Methods: To capture different economic development levels, a multistage stratified cluster sampling method was used. A total of 3 217 children and adolescents aged 12-16 years old from six middle schools and six primary schools in Guiyang, Duyun, Kaili City from Guizhou Province participated in this study. The Achenbach Child Behavior Checklist (CBCL) and a self designed questionnaire were administered between August and December 2021. Results: The overall detection rate of behavioral problems among the students was 16.60% , and the rate was 14.41% and 18.62% among male and female students, respectively. The results of the Logistic regression analysis showed that good peer relationships ( OR=0.25, 0.16, P <0.05) were associated with a lower risk of behavioral problems in children and adolescents. However, the following factors were associated with an elevated risk of behavioral problems: nonparents as the main caregivers( OR = 1.44 ); children s daily screen time was 1-2 h ( OR =1.47) or more than 2 h( OR =2.75); children who had no sports hobbies( OR = 1.36); father parenting for doting or neglect( OR =1.85, 2.01); maternal parenting for doting( OR = 2.12 ); father had bad life habits( OR =1.69); father and mother were occasionally sick( OR =1.30, 1.38)( P <0.05). Conclusion Behavioral problems among children and adolescents in Guizhou Province is related to children s unhealthy living habits and various factors in the family environment. It is suggested that families, schools and relevant functional departments give full play to their own advantages and take comprehensive measures to prevent the occurrence of behavioral problems among children and adolescents.

The coronavirus disease 2019 (COVID-19) pandemic is caused by infection with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is spread primary via respiratory droplets and infects the lungs. Currently widely used cell lines and animals are unable to accurately mimic human physiological conditions because of the abnormal status of cell lines (transformed or cancer cells) and species differences between animals and humans. Organoids are stem cell-derived self-organized three-dimensional culture in vitro and model the physiological conditions of natural organs. Here we showed that SARS-CoV-2 infected and extensively replicated in human embryonic stem cells (hESCs)-derived lung organoids, including airway and alveolar organoids which covered the complete infection and spread route for SARS-CoV-2 within lungs. The infected cells were ciliated, club, and alveolar type 2 (AT2) cells, which were sequentially located from the proximal to the distal airway and terminal alveoli, respectively. Additionally, RNA-seq revealed early cell response to virus infection including an unexpected downregulation of the metabolic processes, especially lipid metabolism, in addition to the well-known upregulation of immune response. Further, Remdesivir and a human neutralizing antibody potently inhibited SARS-CoV-2 replication in lung organoids. Therefore, human lung organoids can serve as a pathophysiological model to investigate the underlying mechanism of SARS-CoV-2 infection and to discover and test therapeutic drugs for COVID-19.
Adenosine Monophosphate/therapeutic use* ; Alanine/therapeutic use* ; Alveolar Epithelial Cells/virology* ; Antibodies, Neutralizing/therapeutic use* ; COVID-19/virology* ; Down-Regulation ; Drug Discovery ; Human Embryonic Stem Cells/metabolism* ; Humans ; Immunity ; Lipid Metabolism ; Lung/virology* ; RNA, Viral/metabolism* ; SARS-CoV-2/physiology* ; Virus Replication/drug effects*

Objective:To explore the influencing factors of false-positive serological reaction of syphilis.Methods:A total of 166 patients with false-positive serological reaction of syphilis (false-positive group), 145 patients diagnosed with early syphilis without treatment (positive control group) and 124 persons undergoing entry physical examination (negative control group) were included from January 2017 to February 2020 in Beijing Tongren Hospital, Capital Medical University. The gender, age and underlying diseases of the three groups were compared. Logistic regression was used to analyze the influencing factors of false-positive serological reaction of syphilis. The efficacies of chemiluminescence immunoassay (CLIA) and toluidine red unheated serum test (TRUST) were compared. Paired t test or chi-square test was used for statistical analysis. Results:In the false-positive group (166 cases), the age of 117 cases were more than 50 years old and 49 cases <50 years old. There were significant differences in age ((53.1±13.8) vs (24.7±2.8), t=22.56, P<0.01), autoimmune disease (36.7%(61/166) vs 6.5%(8/124), χ2=35.93, P<0.01), hepatitis (9.6%(16/166) vs 3.2%(4/124), χ2=4.92, P=0.026) and tumor (6.6%(11/166) vs 0.8%(1/124), χ2=4.68, P=0.030) between the false-positive group and the negative control group. There were significant differences in gender (there were 91(54.8%) males and 75(45.2%) females in the false-positive group, and 103(71.0%) males and 42(29.0%) females in the positive control group, χ2=8.67, P=0.003), age ((53.1±13.8) vs (34.4±12.9), t=20.13, P<0.01) and autoimmune disease (36.7%(61/166) vs 6.9%(10/145), χ2=39.14, P<0.01) between the false-positive group and the positive control group. Multivariate logistic regression analysis showed that gender (odds ratio ( OR)=2.692, 95% confidence interval ( CI) 1.504-4.816, P=0.001), age ≥50 years old ( OR=30.512, 95% CI 15.959-58.335, P<0.01), autoimmune disease ( OR=2.677, 95% CI 1.258-5.695, P=0.011) and hepatitis ( OR=4.408, 95% CI 1.799-10.799, P=0.001) were the influencing factors of false-positive serological reaction of syphilis. In the false-positive group, the positive rate of TRUST was 84.9% (141/166), which was higher than that of CLIA (23.5%(39/166)). The difference was statistically significant ( χ2=126.25, P<0.01). CLIA was 1.0-10.0 cut off index (COI) in 36 patients, and >10.0 COI in three patients.The difference was statistically significant ( χ2=52.51, P<0.01). The titers were ≤1∶4 in 139 patients and≥1∶8 in two patients with TRUST positive.The difference was statistically significant ( χ2=262.35, P<0.01). The sensitivity and specificity of CLIA were 95.2% and 96.0%, respectively, and those of TRUST were 77.2% and 91.1%, respectively. Conclusions:The influencing factors of false-positive serological reaction of syphilis include patients age ≥50 years and with autoimmune disease or hepatitis. The false-positive rate of TRUST is significantly higher than CLIA.

Objective: To explore the effects of Shenqi Yizhi Granule on the cognitive function and the expression of MMP-2 and VEGF in serum of APP/PS1 transgenic mice. Methods: Taking APP/PS1 double transgenic mice as the research carrier, Shenqi Yizi granule was used to interven for 180 days. The learning and remembering ability was tested by Morris water maze, passive avoidance test, and open field. The pathomorphism of the hippocampus CA1 region of mice was tested by HE staining and electron microscopy. The concentration of MMP-2 and VEGF in serum was tested by ELISA. Results: Compared with the blank group, the escape latency of the model group was significantly increased (P ＜ 0.05), and the frequency of the entering medium ring was obviously decreased (P ＜ 0.05). The number of nerve cells in the hippocampal CA1 area was reduced, and the arrangement was disordered. The nuclear pyknosis was deeply stained, and the ultrastructure of nerve cells was abnormal. The concentration of MMP-2 and VEGF in serum was significantly increased (P ＜ 0.05). Compared with the model group, the escape latency of treatment group was decreased (P ＜ 0.05) and the frequency of the entering middle ring was increased (P ＜ 0.05). The concentration of MMP-2 and VEGF in serum was decreased (P ＜ 0.05). Conclusion: Shenqi Yizhi Granule could improve the ability of learning and memory of APP/ PS1 double transgenic mice, and decrease the concentration of MMP-2 and VEGF in serum.

Alzheimer's disease (AD) is a complex neurodegenerative disease. Its pathogenesis involves many factors such as environment, heredity, aging, diet, personal preference and underlying diseases. And the complex pathogenic factors of AD lead to many social and economic problems such as delayed diagnosis, difficulty of drug research and development, low cure rate and high cost of care. In this paper, the relationship between AD disease progression and its risky diseases such as metabolic diseases, chronic inflammatory diseases and cardiovascular diseases was analyzed based on energy metabolism abnormalities. The role of energy metabolism signal/path abnormality activity in the course of risk disease to AD disease was analyzed. Finally, it is suggested that the prevention and treatment of risk disease evolution and adjustment of abnormal energy metabolism signals may be effective strategies for the treatment of AD.

Objective: Based on the Chinese Medicine Heritage Auxiliary Platform (V2.5) software, to analyze the differences in prescriptions for the treatment of Alzheimer's disease and Vascular dementia in the literature of the past five years, and to find the rules of prescription. Methods: Screening CNKI, VIP, and Wan fang data for nearly 5 years to find the prescriptions for the treatment of Alzheimer's disease and Vascular dementia, and constructing a prescription database through the Chinese Medicine Heritage Auxiliary Platform (V2.5) software, and utilizing the Data mining algorithms such as association rules and entropy clustering of the software. Compare the frequency of medication, the drug.s natural, flavor and channel tropism, the common drug pairs, the potential drug pairs, and the differences in the new prescriptions used in the treatment of senile dementia and vascular dementia. Results: 13 of the top 20 high frequency herbs used in AD and VD are the same, the other is different. Its common pathogenesis reflect the interaction of deficiency, stasis and phlegm, while the difference shows that AD focuses on spleen and kidney deficiency, insufficiency of essence and blood, but VD focuses on the interaction of phlegm blood stasis and toxic turbidity obstruction in collaterals. They both use warm herbs in nature and flavor, but AD mainly use sweet and warm herbs, followed by calm and cold herbs, while VD mainly use bitter and warm herbs, followed by cold and calm herbs; The channel tropisms both are mainly liver channel, followed by heart channel, spleen channel and kidney channel. In addition, 16 potential medicine pairs, 7 new prescriptions, 26 new prescriptions and 13 new prescriptions were found. Conclusion:Prescriptions for AD and VD have similarities and differences, with emphasis on each other, suggesting that the former focuses on warming Yang and invigorating spleen, while the latter focuses on resolving phlegm and removing blood stasis, detoxifying and awakening the brain.

It is great value to analyze the pathogenesis of Alzheimer's disease for the formulation and research of prevention and treatment strategies. The change of zang-fu meridians and collaterals is the essential rule of the pathogenesis of internal injury and miscellaneous diseases. Based on the analysis of the physiological basis of the aging changes of zang-fu meridians and the relationship between dementia and the abnormal function of zang-fu meridians and collaterals, this paper systematically explored the relationship between the changes of zang-fu meridians and the pathogenesis of dementia. It is considered that the change of zang-fu meridians and collaterals is the basic law of its pathogenesis development. Therefore, it is proposed that strengthening Qi is the essential principle throughout the whole process of dementia, and dredging channels and collaterals and maintaining normal function of meridians and collaterals are the key to block the pathogenesis evolution of dementia.

Objective To establish a liquid chromatography-tandem mass chromatography ( LC-MS/MS) method for determination of vancomycin concentration in human serum and compare its methodological performance with chemiluminescence microparticle immuno-assay (CMIA). Methods The proteins in serum sample were precipitated by methanol and then vancomycin was eluted and separated on Agilent Poroshell 120EC C18 column (2.1 mm×50 mm, 2.7 μm) with mobile phase of methanol and water in a gradient percentage. Both phases of methanol and water contained 0.1% formic acid. The flow rate was 0.5 mL/min. Norvancomycin was applied as internal standard. Electrospray ionization source was applied and operated in positive ion mode of multiple reaction monitoring (MRM). A total of 112 serum samples collected from the patients taking vancomycin in our hospital, and the results were compared with those of CMIA. Results The method exhibited good linearty within the concentration range of 1 to 100 μg/mL (r2＝ 0.9964).The intra-and inter-day accuracy and precision were all satisfactory for the requirements of quantitative drug testing. No matrix effect and carry-over effect were observed. The results of Wilcoxon symbol rank test showed a difference with statistical significance was found between the serum con-centrations of vancomycin determined by LC-MS/MS and CMIA (P＜0.05), but strong positive correlation and good linear regression were shown between the two methods (Y＝1.06X－0.37, r＝0.986). Conclusion LC-MS/MS should be a cost-saving method superior to CMIA. Its results highly correlated with those of CMIA method. The advantages of LC-MS/MS on accuracy and precision obviously outperformed those of CMIA. Since all the results of LC-MS/MS are comparable to CMIA, this method should be promising to use in determining concentration of vancomycin in serum samples with high clinical value.