ObjectiveThis paper aims to observe the syndrome improvement and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children （heat accumulation in the lung and stomach syndrome）. MethodsThrough a multi-center randomized controlled methodology design，confirmed influenza cases were collected from October 2022 to April 2023 in the pediatrics department of eight hospitals，such as Dongfang Hospital of Beijing University of Chinese Medicine. A total of 180 children with influenza and heat accumulation in the lung and stomach syndrome conforming to the standard were recruited through the clinic. The sick children meeting the inclusion criteria were randomly divided into groups by a block-randomized method. The children in the experimental group were treated with Qingxuan Daozhi formula for five days，and those in the control group were treated with Oseltamivir Phosphate Granules for five days. The primary efficacy indicator was the negative conversion rate of influenza antigen detection. Secondary efficacy indicators were the Canadian acute respiratory illness and flu scale （CARIFS） and the incidence of complications，severe cases， and critical cases. Follow-up observation was conducted on the day of enrollment，48 hours after medication，72 hours after medication， and （6+1） d after medication. ResultsOne hundred and eighty participants were randomly assigned to the experimental group （90 cases） or the control group （90 cases）. All participants were followed up during the study. Comparison of influenza antigen detection results in the primary efficacy indicators showed that the average time of negative influenza antigen conversion in the experimental group was （5.29±1.25） d，and that in the control group was （5.40±1.68） d，without a statistically significant difference. After five days of intervention，52 cases in the experimental group and 51 cases in the control group converted to negative，without a statistically significant difference. CARIFS score results in the secondary efficacy indicators showed that during 72 hours after intervention，there were statistically significant differences between the experimental group and the control group in three dimensions， including headache，muscle soreness， and the need for extra care （P<0.05）. On the （6+1） days after the intervention，the differences in both the experimental group and the control group were statistically significant in 10 dimensions， including sore throat，bad sleep，uncomfortable feeling，poor spirit and fatigue，crying more than usual，the need for extra care，symptom，function，influence on parents，and total score （P<0.05）. The comparison results within the group in the dimensional scores of symptom， function， and influence on parents，as well as the CARIFS total score showed that with the delay of follow-up time，scores of both groups decreased significantly，with a statistically significant difference （P<0.01）. Inter-group comparison results showed that the mean score of the experimental group was higher than that of the control group at the time of enrollment. With the progress of intervention，the score of the experimental group was significantly decreased compared with that of the control group. At the end of follow-up，the mean score of the experimental group was lower than that of the control group，with no statistically significant difference. In terms of the incidence of complications，severe cases， and critical cases， there were no complications，severe cases， and critical cases in the two groups，without a statistically significant difference. ConclusionThe symptom improvement effect and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children （heat accumulation in the lung and stomach syndrome） are not inferior to Oseltamivir Phosphate granules， and children's acceptance is better. It can be more widely used in clinical treatment of influenza in children （heat accumulation in the lung and stomach syndrome）.

Objective:To design protein binders of human epidermal growth factor receptor-2(HER2,also known as ErbB2)using de novo protein design and to preliminarily evaluate their biological effects on tumor cells.Methods:Based on de novo protein design strategy,RosettaDesign and ProteinMPNN algorithms were used to design the binding proteins targeting ErbB2.Concurrently,AlphaFold was applied to assess structural accuracy of the protein binders and their interactions with ErbB2.Subsequently,yeast surface display technology combined with dual-fluorescence high-throughput flow cytometry were performed to screen for protein binders which were capable of specifically binding to ErbB2 specifically.Selected binding proteins were expressed and purified in Eschrichia coli system.Bio-layer interferometry(BLI)was used to validate the binding specificity of the purified protein binders to ErbB2.Finally,Western blotting and CCK-8 assay were used to evaluate the effects of the candidate binding protein on the downstream signaling of ErbB2 and cell proliferation capacity in ovarian cancer(SK-OV-3)and pancreatic cancer(BxPC-3)cell lines,respectively.Results:In this study,the protein binders targeting ErbB2 were successfully designed,a corresponding binding protein library was established,and a protein binder that specifically binds to ErbB2 was screened out from the library.After BLI verification,this protein binder could effectively bind to ErbB2.Further investigation revealed that in ErbB2-high-expressing SK-OV-3 and BxPC-3 cells,this protein binder could effectively inhibit cell proliferation and reduce the phosphorylation level of AKT,a signaling molecule downstream of ErbB2.Conclusion:Based on the strategy of de novo protein design,this study successfully constructed a protein binder that can effectively bind to ErbB2.The binder can effectively inhibit the activation of the downstream signaling pathway mediated by ErbB2 and the proliferation of tumor cells.This indicates the potential application value of de novo designed protein binders targeting ErbB2 in cancer therapy,providing a novel research approach for the field of targeted tumor therapy.

Objective:To design protein binders of human epidermal growth factor receptor-2(HER2,also known as ErbB2)using de novo protein design and to preliminarily evaluate their biological effects on tumor cells.Methods:Based on de novo protein design strategy,RosettaDesign and ProteinMPNN algorithms were used to design the binding proteins targeting ErbB2.Concurrently,AlphaFold was applied to assess structural accuracy of the protein binders and their interactions with ErbB2.Subsequently,yeast surface display technology combined with dual-fluorescence high-throughput flow cytometry were performed to screen for protein binders which were capable of specifically binding to ErbB2 specifically.Selected binding proteins were expressed and purified in Eschrichia coli system.Bio-layer interferometry(BLI)was used to validate the binding specificity of the purified protein binders to ErbB2.Finally,Western blotting and CCK-8 assay were used to evaluate the effects of the candidate binding protein on the downstream signaling of ErbB2 and cell proliferation capacity in ovarian cancer(SK-OV-3)and pancreatic cancer(BxPC-3)cell lines,respectively.Results:In this study,the protein binders targeting ErbB2 were successfully designed,a corresponding binding protein library was established,and a protein binder that specifically binds to ErbB2 was screened out from the library.After BLI verification,this protein binder could effectively bind to ErbB2.Further investigation revealed that in ErbB2-high-expressing SK-OV-3 and BxPC-3 cells,this protein binder could effectively inhibit cell proliferation and reduce the phosphorylation level of AKT,a signaling molecule downstream of ErbB2.Conclusion:Based on the strategy of de novo protein design,this study successfully constructed a protein binder that can effectively bind to ErbB2.The binder can effectively inhibit the activation of the downstream signaling pathway mediated by ErbB2 and the proliferation of tumor cells.This indicates the potential application value of de novo designed protein binders targeting ErbB2 in cancer therapy,providing a novel research approach for the field of targeted tumor therapy.

Objective:To investigate the effect of de novo designed epidermal growth factor receptor(EGFR)binding protein EGn and fibroblast growth factor receptor(FGFR)Ⅲc binding protein FG2 on pancreatic cancer cells.Methods:EGn and FG2 sequences were obtained from an existing study,and the proteins were amplified and expressed in Escherichia coli BL21(DE3)followed by purification through Ni-NTA affinity chromatography and size exclusion chromatography.The purified protein was tested with SDS-PAGE followed by Coomassie Brilliant Blue staining for purity evaluation.The expression level of EGFR and FGFR2 Ⅲc in PANC-1,MIA PaCa-2 and BxPC-3 cells was compared using Western blotting and real-time fluorescence quantitative PCR.The binding of EGn and FG2 in different pancreatic cancer cell lines was assessed by flow cytometry.The effect of EGn and FG2 on the phosphorylation of the target receptors or the down-stream extracellular signal-regulated kinase(ERK)molecules was analyzed by Western blotting.The effect of EGn and FG2 on the proliferation of PANC-1 and MIA PaCa-2 cells was evaluated by CCK-8 assay and colony formation assay.The effect of EGn and FG2 on the migration of PANC-1 cells was examined by Transwell assay.Results:EGn and FG2 proteins were successfully purified and can specifically bind to pancreatic cancer cell lines expressing target receptors,with the binding efficiency positively correlated with target receptor expression levels.EGn and FG2 can competitively inhibit the down-stream signaling of target receptors in PANC-1 cells,and significantly suppress the proliferation and migration of PANC-1 cells(P<0.05).However,their inhibitory effects on the down-stream signaling or proliferation in MIA PaCa-1 and BxPC-3 cells were relatively limited.Conclusion:EGn and FG2 demonstrate inhibitory effects on the proliferation and migration of PANC-1 cells with high expression of EGFR and ectopic high-expression of FGFR2 Ⅲc,indicating their potential therapeutic efficacy in pancreatic cancer and supporting the potential application of de novo designed binding proteins in targeted therapy for pancreatic cancer.

Objective:To investigate the effect of de novo designed epidermal growth factor receptor(EGFR)binding protein EGn and fibroblast growth factor receptor(FGFR)Ⅲc binding protein FG2 on pancreatic cancer cells.Methods:EGn and FG2 sequences were obtained from an existing study,and the proteins were amplified and expressed in Escherichia coli BL21(DE3)followed by purification through Ni-NTA affinity chromatography and size exclusion chromatography.The purified protein was tested with SDS-PAGE followed by Coomassie Brilliant Blue staining for purity evaluation.The expression level of EGFR and FGFR2 Ⅲc in PANC-1,MIA PaCa-2 and BxPC-3 cells was compared using Western blotting and real-time fluorescence quantitative PCR.The binding of EGn and FG2 in different pancreatic cancer cell lines was assessed by flow cytometry.The effect of EGn and FG2 on the phosphorylation of the target receptors or the down-stream extracellular signal-regulated kinase(ERK)molecules was analyzed by Western blotting.The effect of EGn and FG2 on the proliferation of PANC-1 and MIA PaCa-2 cells was evaluated by CCK-8 assay and colony formation assay.The effect of EGn and FG2 on the migration of PANC-1 cells was examined by Transwell assay.Results:EGn and FG2 proteins were successfully purified and can specifically bind to pancreatic cancer cell lines expressing target receptors,with the binding efficiency positively correlated with target receptor expression levels.EGn and FG2 can competitively inhibit the down-stream signaling of target receptors in PANC-1 cells,and significantly suppress the proliferation and migration of PANC-1 cells(P<0.05).However,their inhibitory effects on the down-stream signaling or proliferation in MIA PaCa-1 and BxPC-3 cells were relatively limited.Conclusion:EGn and FG2 demonstrate inhibitory effects on the proliferation and migration of PANC-1 cells with high expression of EGFR and ectopic high-expression of FGFR2 Ⅲc,indicating their potential therapeutic efficacy in pancreatic cancer and supporting the potential application of de novo designed binding proteins in targeted therapy for pancreatic cancer.

Kidney cancer usually requires multidisciplinary individualized treatments. No matter what kind of treatment, drugs are essential. According to the "six-step process" (prescription legitimacy review, patient basic information evaluation review, treatment protocol review, organ function and laboratory index review, pretreatment review, and unconventional prescription review) in prescription review proposed by the anti-tumor drug prescription review expert group and referring to domestic and foreign kidney cancer guidelines and drug instructions in recent years, this consensus selects 9 targeted drugs and 4 immunotherapeutic drugs that are currently commonly used in China and elaborates the key review points in patient basic information evaluation review, treatment protocol review, and organ function and laboratory index review of kidney cancer drug treatment, in order to provide reference for clinical front-line pharmacists to review prescriptions of kidney cancer patients and promote rational drug use in clinic.

Kidney cancer usually requires multidisciplinary individualized treatments. No matter what kind of treatment, drugs are essential. According to the "six-step process" (prescription legitimacy review, patient basic information evaluation review, treatment protocol review, organ function and laboratory index review, pretreatment review, and unconventional prescription review) in prescription review proposed by the anti-tumor drug prescription review expert group and referring to domestic and foreign kidney cancer guidelines and drug instructions in recent years, this consensus selects 9 targeted drugs and 4 immunotherapeutic drugs that are currently commonly used in China and elaborates the key review points in patient basic information evaluation review, treatment protocol review, and organ function and laboratory index review of kidney cancer drug treatment, in order to provide reference for clinical front-line pharmacists to review prescriptions of kidney cancer patients and promote rational drug use in clinic.

Objective:To explore the pathological characteristics and outcomes of elderly patients with community acquired pneumonia (CAP) accompanied by coronavirus disease 2019 (COVID-19).Methods:The diagnosis and treatment process of one elderly patient with CAP accompanied by COVID-19 who was admitted to COVID-19 Treatment Center of Liaoning Province on February 7, 2020 were reviewed. The experience of treatment by analyzing the characteristics of such type of patients during diagnosis and treatment were summarized.Results:A female patient, aged 79 years ald, was admitted to the Center with following features: fever, dry cough, fatigue with dyspnea, scattered moist rales in both lungs, oxygenation index (PaCO 2/FiO 2) of 95 mmHg (1 mmHg = 0.133 kPa), and diffuse interstitial pneumonia in both lungs indicated by chest CT, of which the majority were ground glass-like and fibrous lesions. It was confirmed to be consistent with the feature of severe COVID-19 cases. The patient was successfully cured one month later following anti-inflammatory, anti-viral and high-flow oxygen therapies, homeostasis maintenance of the body, psychological counseling, etc. Accordingly, the treatment experience in CAP combined with COVID-19 in the elderly patients was summarized as follows. In respiratory system, the timing of high-flow oxygen therapy and mechanical ventilation should be seized. As for anti-inflammatory and antiviral therapy, attention should be paid to the treatment of CAP as well as antiviral therapy and symptomatic and supportive therapy. With the progression of the disease, the production of drug-resistant bacteria and the possibility of fungal infection should be paid attention to. For the circulatory system, we should pay attention to the stability of fluid volume and internal environment, and strengthen hemodynamic monitoring and bedside ultrasound to evaluate the cardiovascular capacity-load. In the aspect of the immune system, the selection of the application time of immune-enhancers and glucocorticoids should be paid attention to. In terms of enteral nutrition, early low-fat and high-protein diet is conducive to the recovery of intestinal function and the prevention of bacterial translocation. In addition to the protection of the function of important organs, therapies such as reasonable sedation and psychological intervention should also be used. Conclusions:Elderly patients with CAP accompanied by COVID-19 have complicated conditions and high degree of difficulty in treatment. Comprehensive evaluation of the disease as well as synthetic and effective intervention are the key factors of successful treatment of such patients.

The application of simulated therapy in surgical clinical teaching can stimulate the students' learning enthusiasm and improve their unified qualities in all-rounded way,improve the teacher's teaching level to acquire satisfactory results.

A comparison teaching experiment was carried out in grade 2001 and grade 2002.The students majoring in clinical medicine in these two grades were taught surgery with graphical approach and traditional method respectively.The student number,the test marks and the survey by questionnaire show that graphical teaching approach is more attractive with its visualization and originality it can stimulate students' interests in learning,develop their thinking and improve their clinical analytic ability.It is an important approach to improve clinical teaching quality to lay a good foundation for students before their clinical practice.