Objective To construct a eukaryotic expression vector for the SARS-CoV-2 Omicron BA.1 mutant S1 protein,ex-pressed in CHO-K1 cells to evaluate its immunogenicity,and investigate the adjuvant and antigen dosages.Methods The recombinant plasmid UCOE-Omi-S1 was constructed and transfected into CHO-K1 engineering cells for expression and purification.The recombi-nant S1 protein was confirmed using SDS-PAGE and Western blot.BALB/c mice were randomly divided into 12groups:PBS group,no adjuvant group(high,medium and low-dose groups),aluminum salt adjuvant control group,MF59 adjuvant control group,aluminum salt adjuvant experimental group(high,medium and low-dose groups),MF59 adjuvant experimental group(high,medium and low-dose groups).The solution in accordance with the required proportion of the group was injected intramuscular into the mice 3 times,the interval of 14days,and the blood was collected from the tail vein every 2 weeks,and the serum was separated 30days after the last immu-nization.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum antibody levels and to neutralize pseudoviruses with SARS-CoV-2 prototype and SARS-CoV-2 Omicron BA.1 variant strains.Results After the expression of the target protein in CHO-K1 engineering cells,the expressed protein was observed at a relative molecular mass of approximately 70kDa and was identified as Omi-S1 protein by Western blot.The immune induction effect of the aluminum salt adjuvant group and MF59 adjuvant group was better than that of the no adjuvant group.There was no significant difference between the aluminum salt adjuvant group and the MF59 adjuvant group,but the MF59 adjuvant took effect more quickly.The immune induction effect of the high,middle,and low-dose groups was simi-lar at the 8th week,but the effect of the high-dose group was faster.Pseudovirus neutralization experiments showed that MF59 adjuvant experimental group exhibited higher levels of neutralizing antibodies against the Omicron variants than the aluminum salt adjuvant group.Conclusion This study successfully constructed a recombinant protein of Omicron S1 with good immunogenicity and generated a strong humoral immune response in mice,inducing high levels of neutralizing antibodies against SARS-CoV-2 pseudovirus.The adjuvant and antigen dosages of SARS-CoV-2 were preliminarily studied showed that a dose of less than 10μg of antigen dosages with MF59 adjuvant was found to be effective in inducing good immunity.These findings provide the experimental basis for the development of recombinant protein vaccines against SARS-CoV-2 variants.

ObjectiveTo explore the antidepressant effect of Sophora flavescens seed extract and its molecular mechanism. MethodA mouse depression model was established by intraperitoneal injection of lipopolysaccharide（LPS）， and normal group， model group， fluoxetine group（2.5 mg·kg^-1）， and S. flavescens seed low， medium and high dose groups（200， 400， 800 mg·kg^-1） were set up for 7 d of consecutive gavage. Then the antidepressant effect of S. flavescens seed extract was evaluated by using open field test， elevated plus maze test and forced swimming test. Pathological morphological changes in the hippocampal tissue was observed by hematoxylin-eosin（HE） staining. Protein expression levels of G₁/S-specific cyclin D₁（Cyclin D₁）， Wnt1， β-catenin and phosphorylated glycogen synthase kinase-3β（p-GSK-3β） in mouse brain tissues were detected by Western blot. Hippocampal cell apoptosis was detected by terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate（dUTP） nick end labeling（TUNEL）. ResultThe results of mouse behavioral experiments showed that compared with the normal group， the speed of movement in the open field and the distance of movement in the central area of the open field， and the time spent on the open arms of the elevated plus maze were significantly reduced in the model group（P<0.01）， while immobility time in the forced swimming test was significantly increased（P<0.05）. Compared with the model group， the S. flavescens seed medium and high dose groups had increased speed of movement in the open field test and time spent on the open arms of the elevated plus maze test（P<0.05， P<0.01）， and decreased immobility time in the forced swimming test（P<0.05）， the distance of movement in the central area of the open field test increased in the high dose group（P<0.05）. HE staining results showed that compared with the normal group， the hippocampal neuron structure of mice in the model group was damaged. Compared with the model group， after treatment of S. flavescens seed extract， the pathological state of the mouse hippocampal neuron structure was alleviated， and the neurons increased， were neatly arranged， and the cytoplasm was clear. Western blot results showed that the protein expression levels of Wnt1 and β-catenin in mouse brain tissue were significantly decreased（P<0.01）， while the protein expression levels of Cyclin D₁ and p-GSK-3β were significantly increased（P<0.01） after LPS injection. Compared with the model group， protein expression levels of Wnt1 and β-catenin in brain tissue of S. flavescens seed medium and high dose groups were significantly increased（P<0.01）， while the protein expression levels of Cyclin D₁ and p-GSK-3β were significantly decreased（P<0.01）. TUNEL staining results showed that the hippocampal cell apoptosis rate in the model group was significantly increased compared with that of the normal group（P<0.01）， while the hippocampal cell apoptosis rate in the S. flavescens seed medium and high dose groups was significantly decreased compared with that of the model group（P<0.01）. ConclusionS. flavescens seed extract can effectively improve the severity of depression in LPS-induced depressed mice， and its molecular mechanism is related to the regulation of neuroinflammation and hippocampal neuronal apoptosis mediated by Wnt/β-catenin signaling pathway.

BACKGROUND: Among malignant tumors, lung cancer has the highest mortality rate. Small cell lung cancer (SCLC) is a kind of malignant lung cancer. Its doubling time is very fast. Patients are prone to drug resistance during treatment, and their condition often deteriorates rapidly after recurrence. Except for topotecan, there is a lack of effective second-line single-agent chemotherapy. This study aims to analysis the efficacy and safety of irinotecan (CPT-11) in the second-line treatment of refractory and relapsed SCLC. METHODS: A total of 107 SCLC patients were collected from the Department of Oncology, Jilin Guowen Hospital, who were diagnosed from April 2012 to March 2020, relapsed within 6 months after first-line treatment, and received second-line chemotherapy with single-agent CPT-11. Follow-up until November 2020, calculate the patient's progression free survival (PFS) and overall survival (OS), and summarize the effects and adverse reactions of CPT-11 chemotherapy. RESULTS: The patient's median PFS was 3.8 (3.4-4.4) months, median OS was 8.1 (6.5-10.9) months, objective response rate (ORR) was 16.82% (18/107), and DCR was 55.14% (59/107). The incidence of grade 3-4 adverse reactions in patients was relatively low. Among them, neutropenia was 13.08%, delayed diarrhea was 7.48%, nausea and vomiting was 17.76%, and liver function impairment was 6.54%. The influencing factors of PFS in single-agent CPT-11 second-line chemotherapy were gender (P=0.001), NSE (P=0.029), and effusion (P=0.040). While the influencing factors of OS were NSE level only (P=0.033). CONCLUSIONS For patients with refractory relapsed SCLC, CPT-11 single-agent second-line chemotherapy has a certain effect, is well tolerated, and is worthy of promotion.
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Objective:To explore the clinical utility of liquid chromatography tandem mass spectrometry forprimary aldosteronism screening.Methods:From January to October 2019, 413 inpatients diagnosed hypertension from Fuwai Hospital of Chinese Academy of Medical Sciences were enrolled, including 60 Primary aldosteronism(PA)patients and 353 primary hypertension patients. The plasma aldosterone concentration (PAC) and renin concentration (DRC) were measured after 2 h of standing. The 24 h urine samples were collected for measurement of aldosterone using LC-MS/MS. The performance of urine aldosterone and urine aldosterone/renin ratio (UADRR) in PA screening was evaluated by ROC, and compared with PAC/DRC ratio (ADRR). Meanwhile, the efficiency of urine aldosterone in elderly patients or patients with low blood potassium or 24 h urine sodium over 200 mmol was investigated.Results:Area under the curve (AUC)of urine aldosterone was 0.725 (95 %CI 0.679-0.767), and the best cut-off was 7.13 μg/24 h, which was lower than AUC of ADRR (0.958, 95 %CI 0.934-0.975). The AUC of UADRR was 0.947 (95 %CI 0.920-0.966), the best cut-off was 1.11 (μg/24 h)/(μIU/ml), the sensitivity and specificity were 91.7% and 89.0%, respectively. There is no significant differences found with ADRR. In patients with 24 h urine sodium over 200 mmol, AUC of aldosterone was 0.834 (95 %CI 0.730-0.910) and the best cut-off was 9.31 μg/24 h. The sensitivity and specificity were 90.9% and 68.7%, respectively. For the elderly patients over 60 years old, the AUC of urinary aldosterone was 0.860 (95 %CI 0.770-0.925), and the best cut-off was 6.91 μg/24 h. The sensitivity and specificity were 84.6% and 81.3%, respectively. When admission blood potassium was less than 3.50 mmol/L, AUC of urinary aldosterone was 0.822 (95 %CI 0.684-0.917), and the best cut-off was 10.63 μg/24 h. The sensitivity and specificity were 85.7% and 66.7%, respectively. Conclusion:The detection of aldosterone in urine by LC-MS/MS can provide clinical information for PA screening, and the screening performance is better in patients with 24-hour urine sodium over 200 mmol, elderly patients or patients with low blood potassium. If combined with renin, screening efficiency was the same as that in ADRR.

OBJECTIVE： To investigate the effects of ligustrazine structural modification product Liguzinediol on hemodynamics in chronic heart failure （CHF） model rats induced by adriamycin. METHODS： SD rats were given intraperitoneal injection of adriamycin （2 mg/kg） to induce CHF model. Model rats were randomly divided into normal saline group， positive control group （Deacetyl tricyanidin injection， 0.022 5 mg/kg） and Liguzinediol low-dose， medium-dose and high-dose groups （5， 10， 20        mg/kg）， with 8 rats in each group. Other 8 normal rats were selected as blank control group （normal saline）. Each group was given relevant medicine intravenously. The left ventricular systolic pressure （LVSP）， maximal rate of rise or drop of left ventricular     （±dp/dtmax）， systolic pressure （SP）， diastolic pressure （DP）， heart rate （HR） and other hemodynamic indexes were recorded by multichannel physiological recorder at 1， 5， 10， 20， 40， 60， 90， 120 min after medication. RESULTS： Compared with blank control group， LVSP， +dp/dtmax， │－dp/dtmax│， SP， HR and DP at 120 min after medication of normal saline group were decreased significantly （P＜0.05）. Compared with normal saline group， LVSP at 5-60 min after medication， +dp/dtmax at 40-90 min after medication， SP at 10-40 min after medication were increased significantly in Liguzinediol low-dose group （P＜0.05 or P＜0.01）. LVSP at 5-90 min after medication， SP at 10-60 min after medication， DP at 10-60 min （except for 20 min） after medication were increased significantly in Liguzinediol medium-dose group （P＜0.05 or P＜0.01）. LVSP at 1-120 min after medication， +dp/dtmax at 5-90 min after medication， │－dp/dtmax│， and SP at 5-60 min after medication， DP at 40-60 min after medication were increased significantly in Liguzinediol high-dose group （P＜0.05 or P＜0.01）. CONCLUSIONS： Single intravenous injection of Liguzinediol can significantly enhance ventricular systolic function of CHF model rats so as to control or relieve CHF.

Objective: To understand the status of awarences and the influencing factors of food safety among medical students in Shangdong Province,and to provide a reference for improving a healthy eating habit among students on their knowledge about food safety,attitude and behavior. Methods: A total of 2 322 students from 2 medical colleges and universities in Shandong province selected through stratified cluster sampling were investigated with questionnaires. Results: Univariate analysis of variance showed that food safety knowledge differed by gender, grade, major, origin of student, whether learned nutrition knowledge, monthly cost on food (χ2/H=20.48, 128.02, 98.61, 36.50, 77.60, 171.03,P<0.01). In multiple Logistic regression analysis, results showed that gender, major,origin of student, monthly cost on food and the attention of food affect college students’ food safety awareness (P<0.05). Conclusion Food safety awareness among medical students in Shandong Province is relatively high but varies by multiple factors. It is necessary to improve food safety awareness of medical students through various channels.

Objective To investigate the effect of flipped classroom based on WeChat and mi-crolecture in medical chemistry experiment course in the context of "internet". Methods The classes were randomly divided into 2 groups, experimental group (flipped classroom teaching, n=97) and the control group (traditional teaching, n=98). Comparison of the chemistry experiment test results were performed with the use of t test between the two teaching groups at the end of the semester to evaluate the experimental teaching method. All statistical processing and analyses were performed with SPSS software (version 12.0). Results The chemistry experiment test score of the experimental group was higher than that of the control group [(77.84±8.22) vs. (73.43±10.14), t=3.341, P=0.008), and the difference was statistically significant. The results of the questionnaire showed that the students in the experimental group generally consider that flipped classroom teaching is better than the traditional teaching in terms of the cultivation of comprehen-sive quality and the teaching effect. Conclusion In the context of "internet", flipped classroom teaching with WeChat microlecture can better mobilize the enthusiasm of students to learn and participate in medical chemistry experiment course, which has been welcomed by students and further suggest good application prospects.

AIM To explore the effects of Sangtong alkaloids (total alkaloids and total flavones from Mori folium,STA) on the random blood glucose,starch tolerance and hepatic insulin resistance in db/db mice with type 2 diabetes mellitus.METHODS Eight-week-old db/db mice were divided into model group (normal saline),acarbose group (39 mg/kg) and Sangtong alkaloids groups (105,210 and 420 mg/kg),db/m mice were used as control group (normal saline).The mice were given by intragastric administration for one hundred days.The random blood glucose of mice was determined every ten days.The starch tolerance was determined in the 100th day,together with the determination of serum insulin level,insulin resistance index and insulin sensitivity index.Histopathology changes of pancreas were observed by HE staining.Protein expressions of P-IRS1,P-PI3 K,P-AKT and GLUT2 were detected by Western blot.RESULTS Sangtong alkaloids significantly decreased the random blood glucose,serum insulin level and insulin resistance index,and increased the insulin sensitivity index in db/db mice.Meanwhile,Sangtong alkaloids ameliorated the pancreas histopathological damage and up-regulated the protein expressions of P-IRS1,P-PI3K,P-AKT and GLUT2 in liver.CONCLUSION Sangtong alkaloids can decrease the random blood glucose and improve the insulin resistance of liver in db/db mice with type 2 diabetes mellitus,whose mechanism may be associated with the regulation of hepatic insulin signal pathway.

Heat shock protein 27 ( HSP27 ) is an endogenous protein that plays an important role in a great variety of physio-logical and pathological processes. It can express a large number under body stress conditions. Recent studies have shown estro-gen upregulates the expression of HSP27 through a number of ways, playing a perfect “triple protection” role. In the early stage of atherosclerosis, estrogen induces the phosphorylation of HSP27 via PI3K/Akt signaling pathway. Phosphorylation of HSP27 can resist the injury of vascular endothelial cells( VECs) through an antioxidant and anti-apoptotic pathway as well as the inhibition of cytochrome C. In the stage of forming foam cells, estrogen induces the expression and release of HSP27 from mac-rophages by stimulating the estrogen receptor β ( ERβ) , then HSP27 inhibits the LDL uptake and the release of proinflammato-ry cytokine by binding scavenger receptor A ( SR-A) . During the proliferation and migration of vascular smooth muscle cells ( VSMCs) , estrogen induces estrogen receptor α ( ERα) and protein phosphatase 2 ( PP2A) to form a complex that enhances the activity of PP2A, then it can lead to the dephosphorylation of HSP27 and finally inhibit cells proliferation and migration. In summary, the anti-atherosclerotic effect of estrogen is closely re-lated to the role of HSP27. Given the side effects of estrogen re-placement therapy( MHT) , regulating HSP27 may provide a no-vel therapy for the prevention and treatment of cardiovascular dis-eases in menopausal women clinically.

Cardiovascular disease is a serious threat to human health and quality of life ,and it has become the leading cause of death in human.Thus,looking for effective drugs to reduce the mortality and morbidity of such a disease has become a problem to be solved.Due to its good efficacy of activating blood circula-tion and dissipating blood stasis,salvia miltiorrhiza has been widely used in the treatment of cardiovascular disease and ob-tained a good curative effect.Salvianolic acid B is one of the main water-soluble components of salvia miltiorrhiza extract and studies have shown that salvianolic acid B possesses many biolog-ical activities,which not only has a good protective effect on my-ocardial infarction,but could significantly alleviate myocardial ischemia-reperfusion injury.This article reviews the research progress of salvianolic acid B in cardiovascular diseases,and al-so includes discussion about the mechanisms of salvianolic acid B in the regulation of cardiovascular diseases,which may provide references for follow-up research and clinical treatment.