Objective:To explore the needs and feelings of emergency palliative care from the perspective of medical and nursing staff.Methods:Purposeful sampling was used to recruit 23 emergency medical and nursing staff with experience in end-of-life care from two Class Ⅲ Grade A hospitals in Wenzhou City between September and October 2024. Semi-structured interviews were conducted with the respondents. Results were analyzed using NVivo 12 software.Results:Emergency patients at the end of life and their families had immediate needs, including acute symptom control, pain management, and respiratory distress treatment needs, urgent decision support needs, cultural conflict needs, immediate emotional comfort needs and alexithymia. Medical and nursing staff faced numerous obstacles when providing palliative care in emergency settings, such as role conflicts and skill gaps among emergency medical and nursing staff, lack of dedicated palliative care areas and noisy environments that hindered communication due to physical space limitations in the Emergency Department, short decision-making time and difficulties in referral due to limited emergency room time window, the multidisciplinary collaboration in the emergency department lags behind and the response of the specialized team was delayed, and lack of information sharing between emergency departments and wards.Conclusions:Emergency palliative care is characterized by both urgent needs and implementation barriers. It is necessary to optimize the physical space of the Emergency Department, establish a rapid decision mechanism for medical and nursing staff, strengthen multidisciplinary collaboration, and conduct professional training to build a palliative care model suitable for emergency situations.

Objective:To explore the needs and feelings of emergency palliative care from the perspective of medical and nursing staff.Methods:Purposeful sampling was used to recruit 23 emergency medical and nursing staff with experience in end-of-life care from two Class Ⅲ Grade A hospitals in Wenzhou City between September and October 2024. Semi-structured interviews were conducted with the respondents. Results were analyzed using NVivo 12 software.Results:Emergency patients at the end of life and their families had immediate needs, including acute symptom control, pain management, and respiratory distress treatment needs, urgent decision support needs, cultural conflict needs, immediate emotional comfort needs and alexithymia. Medical and nursing staff faced numerous obstacles when providing palliative care in emergency settings, such as role conflicts and skill gaps among emergency medical and nursing staff, lack of dedicated palliative care areas and noisy environments that hindered communication due to physical space limitations in the Emergency Department, short decision-making time and difficulties in referral due to limited emergency room time window, the multidisciplinary collaboration in the emergency department lags behind and the response of the specialized team was delayed, and lack of information sharing between emergency departments and wards.Conclusions:Emergency palliative care is characterized by both urgent needs and implementation barriers. It is necessary to optimize the physical space of the Emergency Department, establish a rapid decision mechanism for medical and nursing staff, strengthen multidisciplinary collaboration, and conduct professional training to build a palliative care model suitable for emergency situations.

Inducing the degradation of KRAS represents a novel strategy to combat cancers with KRAS mutation. In this study, we identify ubiquitin-specific protease 2 (USP2) as a novel deubiquitinating enzyme of KRAS in multiple myeloma (MM). Specifically, we demonstrate that gambogic acid (GA) forms a covalent bond with the cysteine 284 residue of USP2 through an allosteric pocket, inhibiting its deubiquitinating activity. Inactivation or knockdown of USP2 leads to the degradation of KRAS, resulting in the suppression of MM cell proliferation in vitro and in vivo. Conversely, overexpressing USP2 stabilizes KRAS and partially abrogates GA-induced apoptosis in MM cells. Furthermore, elevated USP2 levels may be associated with poorer prognoses in MM patients. These findings highlight the potential of the USP2/KRAS axis as a therapeutic target in MM, suggesting that strategically inducing KRAS degradation via USP2 inhibition could be a promising approach for treating cancers with KRAS mutations.

Objective·To explore the activation methods and activity regulation mechanisms of the testisin zymogen in vitro,laying a foundation for further research on the physiological functions of testisin in organisms.Methods·The eukaryotic expression plasmid for the mouse-derived testisin(mTN)zymogen was constructed through full-gene synthesis and subsequently transfected into eukaryotic HEK293S cells for expression.Purified recombinant testisin protein was obtained through methods such as nickel ion affinity chromatography.Site-directed mutagenesis was performed on the activation site and active center of the zymogen,and the activation mechanism of testisin was analyzed through enzymatic activity assays.Additionally,the activation rate of the protein was determined by altering the incubation conditions of the zymogen.The self-cleavage site of the protease was identified by using Edman degradation and site-directed mutagenesis.Finally,the regulatory mechanism of protease activity was examined by analyzing the proteolytic rate of the protein towards its substrates.Results·The recombinant mTN zymogen was expressed in eukaryotic HEK293S cells,and high-purity,uniform recombinant protein was obtained through a two-step purification process.This protein was found to remain stable under acidic conditions but underwent self-activation in neutral or alkaline environments.The rate of activation was influenced by factors such as pH value and temperature.The self-activation process required on the integrity of the enzyme's activation site,Arg46,and its active center,Ser240.Additionally,the self-activation of the recombinant mTN zymogen was often accompanied by the self-cleavage at the 170/175 loop on the molecular surface,but this self-cleavage did not affect the activity of testisin.Furthermore,the optimal reaction pH for activated mTN was 8.0,with the protein remaining relatively stable near this pH.The optimal reaction temperature was 50℃,while the protein was best stored at temperatures below 30℃.Zn2+and Ca2+were found to significantly inhibit mTN activity.Conclusion·An effective method for the preparation of recombinant mTN zymogen is successfully established,and it is discovered that the recombinant mTN undergoes self-activation and self-cleavage in vitro.

Objective·To prepare a serine protease inhibitor(Serpin)derived from Tannerella which is associated with periodontosis,and analyze its specificity in inhibiting target proteases and its structural characteristics.Methods·Through amino acid sequence analysis,a Serpin from the human oral microbiome database(eHOMD)was selected and expressed in Escherichia coli.The recombinant protein was purified using methods such as nickel ion affinity chromatography.Its specificity in inhibiting serine proteases was analyzed,followed by an analysis of its three-dimensional spatial structure using structural biology methods.Results·A novel Serpin,named Tannerpin-M,with methionine as the active center P1 residue,was identified,and a high-purity recombinant protein was successfully prepared from Escherichia coli BL21(DE3).Further activity testing demonstrated that recombinant Tannerpin-M could effectively form SDS-stable covalent complexes with proteases derived from granulocytes(human neutrophil elastase,cathepsin G,and proteinase 3),as well as with other proteases including kallikrein 1(KLK1),KLK7,and elastase.Tannerpin-M inhibited KLK7 with a second-order association rate constant of 4.12×104 L/(mol·s).The crystal structure of Tannerpin-M in its relaxed state conformation was resolved at a resolution of 2.4 ?(1 ?=0.1 nm).It revealed that Tannerpin-M possessed a significantly elongated reactive center loop and could undergo the classical conformational transition from a stressed to a relaxed state.Conclusion·Tannerpin-M,derived from oral pathogenic bacteria,is a typical inhibitory Serpin,and can effectively inhibit the serine protease released by granulocytes,by which it may protect the oral pathogenic bacteria from attacks of the human immune system.

Objective·To explore the activation methods and activity regulation mechanisms of the testisin zymogen in vitro,laying a foundation for further research on the physiological functions of testisin in organisms.Methods·The eukaryotic expression plasmid for the mouse-derived testisin(mTN)zymogen was constructed through full-gene synthesis and subsequently transfected into eukaryotic HEK293S cells for expression.Purified recombinant testisin protein was obtained through methods such as nickel ion affinity chromatography.Site-directed mutagenesis was performed on the activation site and active center of the zymogen,and the activation mechanism of testisin was analyzed through enzymatic activity assays.Additionally,the activation rate of the protein was determined by altering the incubation conditions of the zymogen.The self-cleavage site of the protease was identified by using Edman degradation and site-directed mutagenesis.Finally,the regulatory mechanism of protease activity was examined by analyzing the proteolytic rate of the protein towards its substrates.Results·The recombinant mTN zymogen was expressed in eukaryotic HEK293S cells,and high-purity,uniform recombinant protein was obtained through a two-step purification process.This protein was found to remain stable under acidic conditions but underwent self-activation in neutral or alkaline environments.The rate of activation was influenced by factors such as pH value and temperature.The self-activation process required on the integrity of the enzyme's activation site,Arg46,and its active center,Ser240.Additionally,the self-activation of the recombinant mTN zymogen was often accompanied by the self-cleavage at the 170/175 loop on the molecular surface,but this self-cleavage did not affect the activity of testisin.Furthermore,the optimal reaction pH for activated mTN was 8.0,with the protein remaining relatively stable near this pH.The optimal reaction temperature was 50℃,while the protein was best stored at temperatures below 30℃.Zn2+and Ca2+were found to significantly inhibit mTN activity.Conclusion·An effective method for the preparation of recombinant mTN zymogen is successfully established,and it is discovered that the recombinant mTN undergoes self-activation and self-cleavage in vitro.

Objective·To prepare a serine protease inhibitor(Serpin)derived from Tannerella which is associated with periodontosis,and analyze its specificity in inhibiting target proteases and its structural characteristics.Methods·Through amino acid sequence analysis,a Serpin from the human oral microbiome database(eHOMD)was selected and expressed in Escherichia coli.The recombinant protein was purified using methods such as nickel ion affinity chromatography.Its specificity in inhibiting serine proteases was analyzed,followed by an analysis of its three-dimensional spatial structure using structural biology methods.Results·A novel Serpin,named Tannerpin-M,with methionine as the active center P1 residue,was identified,and a high-purity recombinant protein was successfully prepared from Escherichia coli BL21(DE3).Further activity testing demonstrated that recombinant Tannerpin-M could effectively form SDS-stable covalent complexes with proteases derived from granulocytes(human neutrophil elastase,cathepsin G,and proteinase 3),as well as with other proteases including kallikrein 1(KLK1),KLK7,and elastase.Tannerpin-M inhibited KLK7 with a second-order association rate constant of 4.12×104 L/(mol·s).The crystal structure of Tannerpin-M in its relaxed state conformation was resolved at a resolution of 2.4 ?(1 ?=0.1 nm).It revealed that Tannerpin-M possessed a significantly elongated reactive center loop and could undergo the classical conformational transition from a stressed to a relaxed state.Conclusion·Tannerpin-M,derived from oral pathogenic bacteria,is a typical inhibitory Serpin,and can effectively inhibit the serine protease released by granulocytes,by which it may protect the oral pathogenic bacteria from attacks of the human immune system.

Objective:To understand the incidence of esophageal cancer in population at high risk in Yangzhong of Jiangsu province.Methods:Cluster random sampling method was conducted to select several natural villages from the high risk area of esophageal cancer in Yangzhong during 2011-2017. Local residents aged 40-69 years were selected as the target population and a total of 14 687 persons were diagnosed esophageal cancer by endoscopy and pathology.Results:Precancerous lesions were detected in 341 persons, with the detection rate as 2.32 % (341/14 687). Among them, there were 234 (1.59 %) cases with mild and 107 (0.73 %) cases with moderate esophageal hyperplasia. There were 77 positive cases with esophageal hyperplasia and the detection rate was 0.52 %. Among these positive cases, 41 showed severe esophageal hyperplasia/carcinoma in situ (0.28 %), with another 19 as early esophageal cancer (0.13 %) and 17 with invasive carcinoma (0.12 %). The overall early detection rate of positivity through active screening was 77.92 % (60/77), with trends of upwarding and then downwarding. The detection rate of esophageal lesions in men was higher than that in women ( P<0.05), but without significant gender specific differences seen in the detection rates of early esophageal cancer or invasive carcinoma ( P>0.05). The detection rates of all lesions other than invasive carcinoma, gradually increased with age ( P<0.05). The detection rate of invasive carcinoma reached the highest (0.39 %, 10/2 547) in the age group of 60-years and slightly decreased in the age group of 65-years old. Conclusions:Considerable numbers of patients with cancer and precancerous lesions in groups at high-risk can be found through the screening program for esophageal cancer, suggesting that the screening program is of great significance in improving the survival rate and quality of life. Attention should be paid to men, with elderly groups in particular, at high-risk in order to increase the detection rate of early cases.

Objective: To evaluate the effectiveness and benefit of the upper gastrointestinal cancer screening in Yangzhong city, Jiangsu province, from 2009 to 2015. Methods: From 2009 to 2015, 31 natural villages with high-incidence of upper gastrointestinal cancer were selected from Baqiao town, Youfang town and Xinglong sub-district in Yangzhong city. 13 776 residents aged 40 to 69 years old were recruited and screened for upper gastrointestinal cancer by using endoscopic examination and pathological diagnosis. Two economic evaluation methods, cost-effectiveness analysis and cost-benefit analysis, were performed to evaluate the current screening schemes. Results: The mean age of all respondents were (53.60±8.14) years old and the males accounted for 43.64% (6 012). A total of 502 cases of upper gastrointestinal tract lesions were detected, including 100 cases of cancer (62 cases of esophagus, gastric/cardiac early stage cancer, 38 cases of advanced stage cancer), 38 cases of severe esophageal hyperplasia/carcinoma in situ, and 15 cases of high-grade intraepithelial neoplasia in stomach/cardia, the detection rate was 0.73%, 0.28% and 0.11%, respectively; the early diagnosis rate was 75.16% (115/153). The cost of a precancerous lesion, a case diagnosed at the early stage and a positive case identified through the upper gastrointestinal cancer screening in Yangzhong City was 10 037.17, 30 460.64 and 22 895.25 RMB, respectively. The early detection cost index from 2009 to 2015 was 0.52, 0.56, 0.48, 0.48, 0.21, 0.30, and 0.26, respectively. The effectiveness-cost ratio from 2009 to 2015 was 3.41, 2.77, 2.66, 2.58, 4.99, 3.12, and 3.48, respectively. Conclusions The project of early diagnosis and treatment of upper gastrointestinal tract cancer in Yangzhong city has achieved good results and benefits.

Objective To evaluate the effectiveness and benefit of the upper gastrointestinal cancer screening in Yangzhong city, Jiangsu province, from 2009 to 2015. Methods From 2009 to 2015, 31 natural villages with high?incidence of upper gastrointestinal cancer were selected from Baqiao town, Youfang town and Xinglong sub?district in Yangzhong city. 13 776 residents aged 40 to 69 years old were recruited and screened for upper gastrointestinal cancer by using endoscopic examination and pathological diagnosis. Two economic evaluation methods, cost?effectiveness analysis and cost?benefit analysis, were performed to evaluate the current screening schemes. Results The mean age of all respondents were (53.60 ± 8.14) years old and the males accounted for 43.64% (6 012). A total of 502 cases of upper gastrointestinal tract lesions were detected, including 100 cases of cancer (62 cases of esophagus, gastric/cardiac early stage cancer, 38 cases of advanced stage cancer), 38 cases of severe esophageal hyperplasia/carcinoma in situ, and 15 cases of high?grade intraepithelial neoplasia in stomach/cardia, the detection rate was 0.73%, 0.28% and 0.11%, respectively; the early diagnosis rate was 75.16% (115/153). The cost of a precancerous lesion, a case diagnosed at the early stage and a positive case identified through the upper gastrointestinal cancer screening in Yangzhong City was 10 037.17, 30 460.64 and 22 895.25 RMB, respectively. The early detection cost index from 2009 to 2015 was 0.52, 0.56, 0.48, 0.48, 0.21, 0.30, and 0.26, respectively. The effectiveness?cost ratio from 2009 to 2015 was 3.41, 2.77, 2.66, 2.58, 4.99, 3.12, and 3.48, respectively. Conclusions The project of early diagnosis and treatment of upper gastrointestinal tract cancer in Yangzhong city has achieved good results and benefits.