Objective To explore the clinical effect of manipulation treatment combined with auricular point sticking on vertebral artery cervical spondylopathy. Methods 120 patients with vertebral artery cervical spondylopathy were randomly divided into 3 groups, 40 cases in each. The comprehensive group received manipulation treatment and auricular point sticking. The manipulation treatment group received manipulation treatment only and the auricular point sticking group received auricular point sticking only. The peak systolic velocity (Vp), end diastolic peak flow velocity (Vd), mean peak flow velocity (Vm) of the vertebral-basilar artery were checked before and after treatment. Results The clinical cure rates (80%) was higher in the comprehensive group than in the manipulation treatment group (52.5%) and in the auricular point sticking group (47.5%) (P<0.01). The Vp, Vd, Vm in 3 groups improved obviously after treatment (P<0.05), especially in the comprehensive group (P<0.01). Conclusion Manipulation treatment with auricular point sticking is more effective on vertebral artery cervical spondylopathy than simply manipulation treatment or auricular point sticking.

[Objective]To investigate the potential mechanism of Yuquan Capsule in treating diabetic nephropathy(DN)by network pharmacology and in vitro experiments.[Methods]The active ingredients and disease-associated genes of Yuquan Capsule were screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Traditional Chinese Medicine Integrated Database(TCMID),the target genes of DN were screened by Genome Annotation Database Platform(GeneCards),Disease Gene Network(DisGeNET)database and Therapeutic Target Database(TTD).The protein-protein interaction(PPI)network was drawn by STRING 11.5 database,and visualized by Cytoscape 3.7.2 software.The key compounds and core targets were screened by CytoHubba and MCODE plugins of Cytoscape 3.7.2 software.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were performed by R software(ClusterProfiler package),and the molecular docking validation was performed by AutoDock and PyMOL software.[Results]There were 177 Yuquan Capsule against DN,477 active components and 135 targets of Yuquan Capsule against DN.The core targets included serine/threonine kinase 1(AKT1),interleukin-1 beta(IL1B),catalase(CAT),nitric oxide synthase 3(NOS3),leptin(LEP),insulin-like growth factor 1(IGF1)and C-C motif chemokine ligand 2(CCL2).GO enrichment analysis showed 2 557 related items,including 1 415 biological processes,657 cellular components and 48 molecular functions.KEGG pathway analysis showed that the targets of Yuquan Capsule against DN were enriched in 66 pathways,of which cyclic adenosine monophosphate(cAMP)signaling pathway and advanced glycation end products-receptor for advanced glycation end products(AGE-RAGE)signaling pathway in diabetic complications may be the key pathways of Yuquan Capsule in the treatment of DN.Molecular docking results showed that the core target and key compounds could bind well.Experimental verification showed that the protein level of CCL2 decreased in a drugdose-dependent manner,and the protein level of CAT and endothelial nitric oxide synthase(eNOs)increased in a drugdose-dependent manner.[Conclusion]Yuquan Capsule has the characteristics of multiple components and multiple targets in the treatment of DN.The mechanism may be related to the down-regulation of CCL2 and up-regulation of CAT and eNOs protein expression,inhibiting oxidative stress,and regulating the key pathway.

Objective:To analyze the online learning behaviors and learning results of students under different teaching modes with different learning motivations, and to provide a reference for optimizing the development of online teaching.Methods:Samples of students taking the physiology small private online course (SPOC) in 2019 and 2020, and students taking the massive open online courses (MOOC) in the same year were selected in the study to discuss the differences in learning behaviors of students under different teaching modes and learning effects on students with different learning motivations. The online learning behaviors of students were analyzed based on their learning engagement, interactive behavior and learning motivation. SPSS 25.0 was used to analyze the data with statistical methods such as function calculation, frequency statistics, regression analysis, rank sum test, correlation test and chi-square test.Results:The learning participation level ( z=14.36, P<0.001), and the human-machine and interpersonal learning interaction level ( z=-11.70, -16.18, both P<0.001) of SPOC learners in 2020 were higher than those in 2019. The overall interactive level was moderately correlated with performances of students ( r=0.42, 0.52, both P<0.001), and the correlation between interpersonal behavior and grades was much more stronger ( r=0.60, 0.55, both P<0.001). The performance composition of SPOC and MOOC learners in 2019 was completely different ( χ2=857.45, P<0.001). The learning effect of externally motivated students was significantly better than that of internally motivated students ( z=-28.42, P<0.001). Conclusion:The teaching mode adopted by the school can affect the learning effect by influencing the students' online learning engagement and interactive behavior. Besides, students' own learning motivation also plays a key role in their academic performance.

Objective To elucidate the prediction ability of monocyte monolayer assay(MMA)used in hemolytic dis-ease of fetus and newborn(HDFN)caused by IgG anti-M.Methods Plasma from eight pregnant women containing IgG an-ti-M were collected,and were divided into two groups(4 cases with HDFN,with severe clinical symptoms such as fetal hy-drops,and 4 cases without HDFN)according to the clinical outcomes.M antigen positive cells were sensitized with dithioth-reitol(DTT)treated plasma from eight pregnant women respectively.MMA was performed by coincubation with monocytes and sensitized M cells,along with negative and positive control set up.T-test was conducted to compare the difference in phagocytic efficiency between two groups.Results The phagocytic efficiency in group with HDFN were 15.37%,13.05%,9.17%and 24.50%respectively,with the mean value of 15.52%,while the group without HDFN were 8.74%,11.07%,5.12%and 6.23%respectively,with the mean value of 7.79%.There was no significant difference in phagocytic efficiency between two groups(P>0.05).The mean values of both groups were not significantly different from the negative control(P>0.05),but both were significantly lower than positive control(P<0.05).Conclusion The low phagocytic efficiency couldn't convince that the MMA is an effective predictor for the HDFN caused by IgG anti-M,indicating that another mech-anism might be responsible for it rather than monocyte phagocytosis.The assessment of the peak systolic velocity in middle cerebral artery of the fetal should be considered in the management for pregnant women who produce IgG anti-M to estimate the situation of fetal anemia.

Objective To investigate high risk human papillomavirus(HR-HPV)prevalence among married women in Beijing and to study the high risk flactors.nethods During March 2007 to September 2008.a total of 6185 married women sampled from 137 communities in 12 districts were screened bv HR-HPV DNA test and cytogical test.The interview was carried out with unified questionnaires.The databage was set up and twice entered in EpiDam 3.0.After checked up,the data were analyzed in SPSS 15.0.Results (1) The HR-HPV infection rate was 9.89%.The HR-HPV infection rate of the city zone,the suburb and the exurb were 9.34%,10.51% and 9.51% (P>0.05).The HR-HPV infection rate of the native and the oudander were 9.53%,11.30% (P<0.05).(2) The age distribution of HR-HPV infection was that the rate was around 10% among 25 to 44 age groups,which was the highest(11.21%) in 30 to 34 age group;then the rate was descended as the age raising,the rate of 50 to 54 age group was the lowest(7.78%).(3) Multiple logistic regression showed that the related risk factors of HR-HPV infection mainly included 1000 RMB and above of family income per person per month.possessing more than 1 sexual partner of her husband,outlander and hish levels of education.(4) The prevalence of cervical intraepithelial neoplasia(CIN)in HR-HPV positive group wag significantly higher than that in HR-HPV negative group(29.76% vs 3.32%,P<0.01).Conclusions(1)The HR-HPV infection rate among aged 25 to 54 years was 9.9% and there was no significant difference in area distribution.(2)The hish risk population which should strengthen screening was the married bearing-age women with high level of family income,outlander,high levels of education and her husband possessing more than 1 sexual partner.(3)HR-HPV infection is the main risk factor for CIN and cervical cancer.while does not provide a causal relationship with them.The high risk population should be checked regularly to understand the development of HR-HPV infection and CIN incidence.

Objective:To investigate the clinical characteristics of children with invasive pneumococcal di-sease (IPD) and the sensitivity of Streptococcus pneumoniae (SP) isolates to antibacterial drugs, so as to provide the reference for diagnosis and treatment of IPD. Methods:The clinical data of IPD patients in the Children′s Hospital of Chongqing Medical University from January 2014 to December 2018 as well as the drug sensitivity results of SP isolates were retrospectively analyzed.Results:The male to female ratio of 139 patients enrolled was 1.5∶1.0.One hundred and sixteen (83.5%) patients were under 5 years old.Of the 31 patients (22.3%) with underlying diseases, 7 patients (5.1%) had hematological malignancy, 6 patients (4.3%) had congenital heart diseases, and 18 patients (12.9%) were immunosuppressed.The common sources of infection were the respiratory system (59.0%, 82/139 cases) and the central nervous system (28.8%, 40/139 cases). The in-hospital mortality rate among them was 15.8% (22/139 cases). More than 90.0% of the SP isolates were insusceptible to Erythromycin, Tetracycline and Clindamycin, and 74.8% (104/139 cases) of the isolates were insusceptible to Meropenem.The susceptibility rate of SP isolates to Amoxicillin was 69.1% (96/139 cases). The SP isolates were completely susceptible to Vancomycin, Linezolid, Moxifloxacin and Ofloxacin.Meningitis isolates had lower susceptibility rates to Penicillin (10.0% vs. 54.5%, P<0.001) and Cefotaxime (32.5% vs. 74.7%, P<0.001) than those in nonmeningitis isolates. Conclusions:Children under 5 years old with underlying diseases are susceptible to IPD and have a high mortality rate.The SP isolates in children with IPD are highly insusceptible to Erythromycin, Tetracycline, Clindamycin and Meropenem, and completely susceptible to Vancomycin, Linezolid, Moxifloxacin and Ofloxacin.Meningitis isolates are less susceptible to Penicillin and Cefotaxime than non-meningitis isolates.

Objective: To explore the relationship between HCM pathogenic gene mutations and clinical phenotypes by analyzing the prenatal diagnosis and genetic characteristics of a pregnant woman from a family with hypertrophic cardiomyopathy (HCM). Methods: The clinical data of the proband and her family members was collected. The DNA was extracted from the peripheral blood, amniotic fluid cells and cultured amniotic fluid cells of proband. Next generation sequencing (NGS) was utilized for screening pathogenetic loci of the proband. The suspected mutation sequences of HCM pathogenic candidate genes MYH7 and MYBPC3 were directly sequenced after PCR. Pathogenicity prediction of amniotic fluid cells was performed by using genetic data and bioinformatics software, such as Mutation taster, PolyPhen-2 and ANTHEPROT. Results: The sequencing results showed that heterozygous mutations of MYH7 c.1988G＞A (p.Arg663His) and MYBPC3 c.151G＞A (p.Ala51Thr) were found in the proband. The phenotype of her father was normal, and no abnormal mutations were detectable. Her mother also showed normal phenotype but carried MYBPC3 c.151G＞A heterozygous mutation. Only MYH7 c.1988G＞A heterozygous mutation was found in the fetus and no abnormal variation of MYBPC3 was showed. The prediction of mutation effect and analysis of protein structure and function revealed that the two missense mutations could affect the hydrophobicity and antigenicity of the protein. The genetic data demonstrated MYH7 c.1988G＞A was defined as a pathogenic mutation. Conclusion MYH7 c.1988G＞A should be a newly generated pathogenic mutation in the proband, or caused by reproductive chimerism of her parents. MYBPC3 c.151G＞A mutation may promote the occurrence of HCM. Although the fetus only carries MYH7 c.1988G＞A, her phenotype may still display as HCM.

Objective: To explore the genetic basis for a pedigree affected with Marfan syndrome (MFS). Methods: Clinical data of the patients was collected.With genomic DNA extracted from peripheral blood samples, potential mutation was detected by targeted exome sequencing.Candidate variants were validated by Sanger sequencing and bioinformatic analysis. Results: Targeted exome sequencing and Sanger sequencing revealed a missense c. 649T>C(p.Trp217Arg) variant in the exon 7 of FBN1 gene, which was unreported previously.Bioinformatics analysis suggested that the variant can cause amino acid replacement and affect the structure and function of fibrillin-1. Conclusion A novel missense variant of the FBN1 gene was identified, which probably underlies the autosomal dominant MFS in this pedigree.

【Objective】 To establish an experimental method for detecting phagocytosis of sensitized red blood cells in vitro by flow cytometry. 【Methods】 Mononuclear cells were isolated from the peripheral blood of blood donors and cultured in a cell incubator for 1 hour, and then adherent monocytes were isolated and obtained. Di^b-positive red blood cells (RBCs) were labeled with PKH26 and then sensitized with IgG anti-Di^b. The sensitized RBCs were added to monocytes for in vitro phagocytosis assay. Monocytes were labeled with FITC anti-human CD14, then phagocytosis was measured by flow cytometry, and the phagocytic efficiency was calculated. The method was used to detect the phagocytic efficiency of monocytes on human IgG anti-D sensitized RBCs with different titers. 【Results】 The phagocytic efficiency of monocytes was averaged at 5% (1.2%~7.6%, SD 3.30) versus 81% (71.4%~92.7%, SD 8.65) in the negative versus positive control group, respectively. Phagocytic activity of monocytes mediated by anti-D was correlated with the antibody titer. The phagocytosis efficiency was within 10% when the antibody titer was lower than 32 and increased sharply when the titer was between 32 to 128, it entered a plateau and stabilized at 80% at the titer above 256. 【Conclusion】 A detection platform for detecting phagocytosis-sensitized RBCs in vitro by flow cytometry has been successfully established. It can be used to assess the clinical significance of red blood cell allotype or autologous IgG antibodies.

OBJECTIVE：To estab lish a method that can comprehensively and rapidly analyze the chemical compositions of Miao medicine Caesalpinia decapetala，and to providing reference for quality control and pharmacodynamic material basis study of C. decapetala . METHODS ：UPLC-Q-TOF-MS/MS was adopted . The determination was performed on Agilent SB-C 18 column with mobile phase consisted of 0.1％ formic acid solution- 0.1％ formic acid acetonitrile （gradient elution ）at the flow rate of 0.25 mL/min. The column temperature was 30 ℃，and sample size was 2 µL. ESI source was applied in negative and positive scanning ion mode and data collection range of m/z 50-1 500. The capillary voltage was 4.5 kV，the atomizing gas （nitrogen）pressure was 1.2 Bar， the solvent removal gas was nitrogen ，the flow rate of solvent removal gas was 8 L/min and the solvent removal gas temperature was 200 ℃. Data Analysis 4.2 software was adopted to analyze fragment ion information of each peak ，and identify chemica l compositions on the basis of relevant literature and mass spectograms of reference substance. RESULTS ：Under positive ion mode ， 9 chemical compounds were identified ；peak 1，2，3，4，5，6，7，8，9 were catechin ，protohematoxylin B ，epicatechin，ethyl gallate，quercetin，luteolin，3-deoxy-hematoxylin chalcone ， isoliquiritigenin and linoleic acid. Under negative ion mode ， U1812403）， totally 21 peaks were confirmed and 13 compounds were identified；peak 3，4，5，6，7，8，9，10，11，12，13，15， 21 were catechins ， brevifolin carboxylic acid ， proto- hematoxylin B ，epicatechin，ethyl gallate ，epicatechin gallate ， quercetin，resveratrol，hematoxylin，luteolin，3-deoxy-hema- toxylin， isoliquiritigenin， linoleic acid. CONCLUSIONS UPLC-Q-TOF-MS/MS method is established successfully for analysis of chemical compositions in C. decapetala .