This study constructed a LHCGR-CRE-luc-HEK293 transgenic cell line according to the activation of the cAMP signaling pathway after recombinant human chorionic gonadotropin binding to the receptor. The biological activity of recombinant human chorionic gonadotropin was assayed using a luciferase assay system. The relative potency of the samples was calculated using four-parameter model. And the method conditions were optimized to validate the specificity, relative accuracy, precision and linearity of the method. The results showed that there was a quantitative potency relationship of human chorinonic gonadotropin (hCG) in the method and it was in accordance with the four-parameter curve. After optimization, the conditions were determined as hCG dilution concentration of 2.5 μg·mL^-1, dilution ratio of 1∶4, cell number of 10 000-15 000 cells/well, and induction time of 6 h. The method had good specificity, relative accuracy with relative bias ranging from -8.9% to 3.4%, linear regression equation correlation coefficient of 0.996, intermediate precision geometric coefficient of variation ranging from 3.3% to 15.0%, and linearity range of 50% to 200%. This study successfully established and validated a reporter gene method to detect hCG biological activity, which can be used for hCG biological activity assay and quality control.

Objective To explore the potential mechanism of action of paeoniflorin in diabetes mellitus,the related targets and pathways were preliminarily discussed,based on the network pharmacology and molecular docking technology.Methods Analyze the potential targets of paeoniflorin using the Swiss Target Prediction database.Genecards and OMIM databases yielded the genes of diabetes-related illnesses.After taking the intersection of the two,protein-protein interaction network(PPI)was established using STRING and Cytoscape programs to search for key genes with strong correlation and complete gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Use AutoDockTools and Pymol programs to complete protein molecule docking validation.Results The pharmacologically-related study revealed 63 targets associated with paeoniflorin,4 758 genes related to diabetes,and 50 intersection targets.15 key genes including vascular endothelial growth factor A(VEGFA),epidermal growth factor receptor(EGFR),V-Ha-ras harvey(HRAS),V-src sarcoma(SRC)and heat shock protein hs 90-alpha(HSP90AA1)were screened.RAs-associated protein 1,Ras,calcium and other signaling pathways were obtained by KEGG pathway analysis.Molecular docking results showed that paeoniflorin had good binding ability with key genes.Conclusion Paeoniflorin can treat diabetes through multiple targets and pathways,and this mechanism can provide a basis for the application of paeoniflorin in anti diabetes and drug research and development.

AIM To study the chemical constituents from seeds of Canavalia gladiata(Jacq.)DC.,and their estrogen-like activities.METHODS The 75% ethanol extract from the seeds of C.gladiata was isolated and purified by various chromatography,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The estrogen-like activities were evaluated by human breast cancer cells(MCF-7)stably transfected with estrogen receptor reporter gene(ER-Luc).RESULTS Twelve compounds were isolated and identified as biorobin(1),kaempferol 3-O-[O-β-D-apio-D-furanosyl(1→2)-O-[α-L-rhamno-pyranosyl(1 → 6)]-β-D-galactopyranoside(2),kaempferol 3-O-{β-D-glucopyranosyl(1 → 2)-[α-L-rhamonopyranosyl(1→6)]-β-D-galactopyranoside}(3),iso-quercitrin(4),quercetin 3-O-(6″-O-galloyl)-β-D-glucoside(5),isomyricitrin(6),myricetin-3-O-rutinoside(7)(3-ethenylphenyl)-1,2-ethanediol(8),1,3,6-tri-O-galloyl-β-D-glucopyranose(9),breynioside A(10),4-hydroxy-3-methoxyphenol-1-O-β-D-(6′-O-galloyl)glucoside(11),maesopsin-6-O-β-D-glucopyranoside(12).Compounds 1,3-5,8-9 in the concentration of 50 μmol/L,and compounds 2,12 in 100 μmol/L showed estrogen-like activities.CONCLUSION All compounds are isolated from the gene Canavalia for the first time.Compounds 1,3-5,8-9 showed stronger estrogen-like activities than compounds 2 and 12.

Objective:Under the background of "new medical science", to construct a whole-process education path with "education as the driving force - moral cultivation as the core", and to provide a decision-making basis for the reform, innovation, and development of humanistic quality education in medical colleges.Methods:Five-year clinical medical students (freshman to fifth graders) from Qiqihar Medical University were selected as the research objects. The evaluation model and questionnaire of humanistic quality of clinical medical students were constructed by literature analysis. The Questionnaire Star platform was used to investigate and collect data by random sampling of clinical medical undergraduates, and the data results were analyzed by SPSS 26.0 to explore the path of building the whole-process education.Results:The empirical research on the cultivation of humanistic quality of medical students showed that the average humanistic attitude score of medical students was (3.47±0.75) points. From the perspective of gender, the scores of humanistic cognition, attitude, and behavior of male students were 2.81, 3.68, and 3.22 points, respectively, which were all significantly higher than those of female students (2.52, 3.22, and 2.95 points, respectively) ( P<0.05). The data from freshmen to the fifth graders showed that humanistic cognition, attitude, and behavior scores all presented an increasing trend with the grade. Using the "Knowledge, Attitude/Belief, Practice" model, it was found that the cognitive rate of humanistic quality of medical students was relatively ideal, and the awareness rate of medical students in different grades was different. The cross analysis showed that the effect of ideological morality and political literacy education on medical students in medical colleges had a significant cross-relationship with multiple choice questions. Conclusion:The humanistic quality of clinical medical students needs to be improved, and the cultivation of medical humanistic quality should run through the whole process from the first year to the fifth year, so as to improve the comprehensive training of professional ability and promote the all-round development of medical students.

Objective To analyze the monitoring data of Oncomelania hupensis in the national schistosomiasis surveillance sites of China from 2015 to 2019, so as to understand the changes of Oncomelania snail status in the schistosomiasis-endemic areas of China and to provide the scientific evidence for Oncomelania snail control. Methods According to the requirements of National Scheme for Schistosomiasis Surveillance in China (2014 Edition), national schistosomiasis surveillance sites were assigned in all schistosomiasis-endemic counties (cities, districts) and the potential endemic counties (cities, districts) in the Three Gorges Reservoir areas, and Oncomelania snail status was monitored according to different epidemic types. In endemic areas, Oncomelania snail survey was performed by means of systematic sampling and environmental sampling, and the occurrence of frames with Oncomelania snails and the prevalence of Schistosoma japonicum infections in Oncomelania snails were calculated, while in potential endemic areas, the risk of imported Oncomelania snails and Oncomelania snails in floating debris were monitored. Results Oncomelania snail survey was performed covering an area of 116 834.16 hm² in the national schistosomiasis surveillance of China from 2015 to 2019, with 35 007.62 hm² Oncomelania snail habitats identified. A total of 6 908 292 frames were surveyed during the 5-year period, and there were 364 555 frames detected with Oncomelania snails, with a 5.28% mean occurrence of frames with Oncomelania snails. Among 997 508 living Oncomelania snails captured, no S. japonicum infections were detected, and loop-mediated isothermal amplification (LAMP) assay detected 18 positive mixed Oncomelania snail samples. During the period from 2015 to 2019, 147.20 hm² emerging Oncomelania snail habitats were identified, with an overall tendency towards a rise seen in the proportion of emerging Oncomelania snail habitats in plain regions with waterway networks (0.12% to 92.00%), a tendency towards a rise followed by decline seen in marshland and lake regions (0 to 96.72%), and a large fluctuation in hilly regions (0 to 88.49%). A total of 831.10 hm² re-emerging Oncomelania snail habitats were found in the national schistosomiasis surveillance sites of China from 2015 to 2019, with an overall tendency towards a rise seen in the proportion of re-emerging Oncomelania snail habitats in marshland and lake regions (16.05% to 79.66%), an overall tendency towards a decline seen in hilly regions (19.25% to 81.00%), and a minor fluctuation in plain regions with waterway networks (1.10% to 10.14%). During the 5-year period from 2015 to 2019, a total of 48 656 kg floating debris were captured in 4 surveillance sites in the Three Gorges Reservoir areas, and 2 204 snails were found, with no Oncomelania snails identified. Conclusions The areas of Oncomelania snail habitats tended to be stable in the national schistosomiasis surveillance sites of China during the period from 2015 to 2019, however, there was a gradual rise in the area of Oncomelania snail habitats year by year, and LAMP assay identified positive Oncomelania snail samples, suggesting Oncomelania snail control is far from optimistic in China.

Objective:To investigate the current status of coping behaviors among caregivers of children with disabilities in the context of rehabilitation and its influencing factors. Methods:From December, 2019 to January, 2020, and August to September, 2020, a total of 358 caregivers (parents) of children with disabilities were surveyed in Shanghai. They were evaluated in the context of rehabilitation of children with disabilities with Coping Health Inventory for Parents (CHIP). Multiple linear stepwise regression analysis was used to explore the influencing factors. Results:The total score of CHIP was (106.00±23.45) and the average score of CHIP was (2.36±0.52). Among the caregiver factor, mothers (B = 7.607, 95%CI 2.477 to 12.737, P = 0.004), and caregivers who didn't need to care for others (B = 5.758, 95%CI 1.174 to 10.343, P = 0.014) reported higher CHIP score. Among the child factors, higher CHIP score was reported in caregivers who had more partners (B = 2.925, 95%CI 1.233 to 4.618, P < 0.001), participated in public space activities more frequently (B = 3.906, 95%CI 1.743 to 6.068, P < 0.001) and perceived changes in rehabilitation outcomes (B = -16.832, 95%CI -31.471 to -2.193, P = 0.024); CHIP score was lower in thoses of 3 to < 6 years old (B = -7.914, 95%CI -15.157 to -0.671, P = 0.032) and with intellectual disability children (B = -11.153, 95%CI -21.360 to -0.947, P = 0.032). Among the environmental factors, caregivers who perceived more friendly social attitude (B = 2.560, 95%CI 0.615 to 4.505, P = 0.010) reported higher CHIP score. Conclusion:It is proposed to learn the coping needs of caregivers of children with disabilities in the context of rehabilitation. The coping behaviors of caregivers are influenced by multiple factors, such as children's individual, caregivers and environment.

Objective To analyze the surveillance data after schistosomiasis transmission interruption from 1985 to 2018, so as to provide scientific basis for the future prevention and control of schistosomiasis. Methods The data pertaining to snail survey, surveillance of schistosomiasis in humans and livestock, and health education were collected and analyzed in Songjiang District after schistosomiasis transmission interruption from 1985 to 2018. Results From 1985 to 2018, there were 8 townships, 68 villages and 1 462 settings with snails in Songjiang District, covering an area of 96.399 hm². There were 4 townships and 59 villages with snails newly identified in previously schistosomiasis non-endemic areas, with emerging snail habitats of 60.161 hm². A total of 26 148 snails were captured, and no infections were detected. The snail habitats were mainly distributed in the river course, ditches and farmlands, which accounted for 45.28%, 33.17% and 21.55% of total snail habitats, respectively. A total of 245 955 humans were subjected to the serological test, with a positive rate of 0.03%, and 37 humans were subjected to the parasitological test, with 3 positives detected. These three egg-positives were all imported chronic cases with schistosomiasis. A total of 9 109 livestock were detected using the serological test, and no infection was found. Most of the awareness rate of schistosomiasis control knowledge and the rate of correct behavior formation were over 80% in residents and students. Conclusions The achievements of schistosomiasis control are consolidated in Songjiang District; however, there are still risk factors of schistosomiasis transmission, including residual snails, snail diffusion and importation of external source of infection. The future surveillance of schistosomiasis should focus on snails and sources of infection.

Objective: To analyze the clinical effect of traditionaldog-days acupoint application and Magic Acupuncture Patch (Manji) in the prevention and treatment of chronic bronchitis (CB) in remission stage in the past five years, and explore the principle of action and effective stimulation, to provide the evidence for treating CB by acupoint application. Methods: A retrospective study was conducted on 405 patients with CB who met the inclusion criteria. All patients were treated with dog-days acupoint application or Magic Acupuncture Patch between 2013 and 2017. The clinical data of 405 patients were statistically analyzed to compare the prevention and treatment effects of dog-days acupoint application and Magic Acupuncture Patch, and different degrees of stimulation of dog-days acupoint application. Results: Among the dog-days acupoint application groups, the total effective rate was 63.6％ in the light stimulation group, 93.1％ in the moderate stimulation group, and 94.8％ in the strong stimulation group. The differences in the total effective rate between the light stimulation group and the moderate stimulation group, as well as the strong stimulation group, were statistically significant (both P<0.05). There was no significant difference in the total effective rate between the moderate stimulation group and the strong stimulation group (P>0.05). The total effective rate was 83.9％ in the dog-days acupoint application group, versus 45.4％ in the Magic Acupuncture Patch group, and there was a statistically significant difference between the two groups (P<0.05). Conclusion: The efficacy of dog-days acupoint application in the prevention and treatment of CB is better than that of Magic Acupuncture Patch; the degree of stimulation is the basis for the effect of dog-days acupoint application on prevention and treatment of CB, and the moderate and strong stimulations are more appropriate.

Objective Quantitative analysis and comparison of the expression of ribonucleic acid （RNA） from frozen organs and formaldehyde-fixed and paraffin-embedded （FFPE） tissues. Methods Frozen specimens of human brain, myocardium and liver tissues as well as FFPE samples at different postmortem intervals were collected and mass concentration of RNA was extracted and detected. Reverse transcription-quantitative polymerase chain reaction （RT-qPCR） technology was used to analyze the amplification efficiency and relative expression of each RNA marker. Results The mass concentration and integrity of RNA extracted from FFPE samples were relatively low compared with frozen specimens. The amplification efficiency of RNA markers was related with RNA species and the length of amplification products. Among them, glyceraldehyde-3-phosphate dehydrogenase （GAPDH） and β-actin （ACTB） with relatively long amplification products failed to achieve optimal amplification efficiency, whereas 5S ribosomal RNA （5S rRNA） achieved ideal amplification efficiency and showed quite stable expression across various tissues, therefore it was chosen as internal reference marker. The expression quantity of GAPDH and ACTB in frozen specimens with longer postmortem intervals and in FFPE samples with relatively long amplification products was decreased. The expressions of tissue-specific microRNAs （miRNAs）, GAPDH and ACTB with relatively short amplification products had consistency in the same tissues and FFPE samples. Conclusion Through standardizing the RT-qPCR experiment, selecting the appropriate RNA marker and designing primers of appropriate product length, RNA expression levels of FFPE samples can be accurately quantified.
DNA Primers ; Formaldehyde ; Gene Expression Profiling ; Humans ; MicroRNAs/analysis* ; Myocardium ; Paraffin Embedding ; RNA/analysis* ; Real-Time Polymerase Chain Reaction/standards*

Objective To analyze the status quo of patients with severe mental illness in Jinshan district, Shanghai, and provide sci-entific evidence for strengthening the prevention and treatment of them. Methods The data of severe mental illness until March 1st, 2016 were derived from the national database and analysis system, and were used for statistical analysis. Results A total of 4778 patients with severe mental illness were registered. Among them, 54.40% suffered from schizo-phrenia, patients aged 18 to 59 years old (67.73%) accounted for the vast majority, 86.44% only got junior high school education or below, 40.46% were unmarried or divorced, 76.50% patients choose outpatient department as a way to see doctor, and 15.11% patients did not take any treatment. Conclusion The prevention of severe mental illness has become the important gambit for this district. The proportion of patients choosing appropriate way to visit doctor is relatively low. The management of patients with severe men-tal diseases needs to be strengthened to promote the recovery of patients and social stability.