Objective To construct the K64 capsule depolymerase recombinant protein,Dep44,and investigate its potential application against carbapenem-resistant Klebsiella pneumoniae(CRKP)infections.Methods The de-polymerase-encoding phage vB_Kpn_HF1013(GenBank:PP803128)was isolated and genomically analyzed to screen for candidate depolymerases.The recombinant protein Dep44 was constructed and functionally verified for depolymerase activity.Dep44 sensitive range was validated and Dep44 antimicrobial activity was assessed by bio-film disruption and serum sterilization assays.Results The tail spike protein of phage vB_Kpn_HF1013 exhibited depolymerase activity and recombinant protein Dep44 specifically degraded K64 CRKP capsule.Biofilm eradication assays demonstrated that recombinant Dep44 at both 2 μg/mL and 10 μg/mL significantly disrupted bacterial bio-films relative to the control.Serum bactericidal assays showed that Dep44 exhibited synergistic activity with serum,dependent on the complement system,as Dep44 alone lacked bactericidal properties.Conclusion Dep44 effec-tively targets and degrades K64 CRKP capsule,disrupts biofilms,and enhances serum bactericidal activity,high-lighting its potential for managing K64 CRKP infections and clearing biofilms from medical devices.

Objective To construct the K64 capsule depolymerase recombinant protein,Dep44,and investigate its potential application against carbapenem-resistant Klebsiella pneumoniae(CRKP)infections.Methods The de-polymerase-encoding phage vB_Kpn_HF1013(GenBank:PP803128)was isolated and genomically analyzed to screen for candidate depolymerases.The recombinant protein Dep44 was constructed and functionally verified for depolymerase activity.Dep44 sensitive range was validated and Dep44 antimicrobial activity was assessed by bio-film disruption and serum sterilization assays.Results The tail spike protein of phage vB_Kpn_HF1013 exhibited depolymerase activity and recombinant protein Dep44 specifically degraded K64 CRKP capsule.Biofilm eradication assays demonstrated that recombinant Dep44 at both 2 μg/mL and 10 μg/mL significantly disrupted bacterial bio-films relative to the control.Serum bactericidal assays showed that Dep44 exhibited synergistic activity with serum,dependent on the complement system,as Dep44 alone lacked bactericidal properties.Conclusion Dep44 effec-tively targets and degrades K64 CRKP capsule,disrupts biofilms,and enhances serum bactericidal activity,high-lighting its potential for managing K64 CRKP infections and clearing biofilms from medical devices.

Immune checkpoint inhibitors (ICIs) are an important method for cancer treatment today, including programmed cell death receptor 1 (PD-1), programmed cell death ligand 1 (PD-L1), and cytotoxic T lymphocyte antigen 4 (CTLA-4) inhibitors. It has achieved unprecedented benefits in various malignant tumors, but its adverse reactions deserve attention. The enhanced immune response induced by these drugs can lead to immune related adverse events (irAEs), which have a significant impact on drug use and patient prognosis. It can occur in any system throughout the body, and ICI associated acute kidney injury (ICI-AKI) is one of them. The mechanisms include loss of tolerance to autoantigens, reactivation of drug specific effector T cells, and production of kidney specific autoantibodies. The most common pathological manifestation is acute tubulointerstitial nephritis, with only a few cases of ICIs related glomerulonephritis reported. ICI-AKI usually responds well to early corticosteroid treatment, and most patients can achieve complete or partial remission. We provide a review of the epidemiology, pathogenesis, clinical manifestations, diagnostic methods, and management strategies of ICI-AKI, with a focus on exploring its risk factors, reactivation of ICIs, and prognosis. This will help to adopt management measures to reduce the occurrence of ICI-AKI and improve the prognosis of tumor patients.

Objective:To observe the clinical efficacy of Yin-Yang balancing acupuncture plus Tuina(Chinese therapeutic massage)for post-stroke depression(PSD). Methods:A total of 72 PSD patients were randomized into an acupuncture-Tuina group and a Western medicine group using the random number table method,with 36 cases in each group.The Western medicine group was treated with fluoxetine hydrochloride capsules.The acupuncture-Tuina group was treated with Yin-Yang balancing acupuncture plus Tuina therapy.Both groups were treated for 4 weeks.After treatment,the clinical efficacy was compared,and the changes in Hamilton depression scale(HAMD)score,National Institute of Health stroke scale(NIHSS)score,serum neurotransmitter levels,and electroencephalogram(EEG)were observed. Results:The total effective rate of the Western medicine group and the acupuncture-Tuina group was 83.3%and 94.4%,respectively.There was a significant difference in the total effective rate between the two groups(P<0.05).After treatment,the serum levels of 5-hydroxytryptamine(5-HT),norepinephrine(NE),and dopamine(DA)were higher than those in the same group before treatment,and the intra-group differences were all statistically significant(P<0.05).The serum levels of 5-HT,NE,and DA in the acupuncture-Tuina group were higher than those in the Western medicine group,and the differences between the groups were statistically significant(P<0.05).After treatment,the scores of HAMD and NIHSS in both groups decreased,and the intra-group differences were statistically significant(P<0.05).The scores in the acupuncture-Tuina group were lower than those in the Western medicine group,and the differences between the two groups were all statistically significant(P<0.05).After treatment,the EEG diffuse wave[(delta+theta)/(alpha+beta)]ratio(DTABR)and delta/beta ratio(DBR)in both groups decreased,and the intra-group differences were statistically significant(P<0.05).The ratios in the acupuncture-Tuina group were lower than those in the Western medicine group,and the differences between the two groups were statistically significant(P<0.05). Conclusion:Yin-Yang balancing acupuncture plus Tuina therapy has a better efficacy than oral fluoxetine hydrochloride capsules in the treatment of PSD,which can increase the levels of serum 5-HT,DA,and NE,and regulate brain waves.

OBJECTIVE To investigate the mechanism of Mayuan tongbian zhitong decoction on improving slow-transmission constipation（STC）in rats by regulating AMP activated protein kinase （AMPK）/endothelial nitric oxide synthase （eNOS）signaling pathway. METHODS The rats were randomly divided into blank group ，model group ，Mayuan tongbian zhitong decoction low-dose，medium-dose and high-dose groups （6，12，18 mg/kg），with 10 rats in each group. Except for blank group ，other groups were given Compound diphenoxylate suspension to induce STC model. After modeling ，blank group and model group were given normal saline intragastrically ，and Mayuan tongbian zhitong decoction groups were given relevant medicine intragastrically ， once a day ，for consecutive 2 weeks. The number of feces and water content of feces in each group were observed before and after treatment；the carbon powder propulsion rate of rats in each group was calculated ；the pathological structure of colon in each group was observed ；the levels of nitric oxide （NO）and nitric oxide synthase （NOS）in colon tissues of rats in each group were detected；the expressions of AMPK ，eNOS，mammalian target of rapamycin （mTOR），tuberous sclerosis complex 1（Tsc-1）， Tsc-2 and eukaryotic promoter 4E binding protein （4ebp）were also detected. The active ingredients of Cannabis sativa ，Citrus aurantium and Rehmannia glutinosa were screened from Mayuan tongbian zhitong decoction. The active ingredients with high Degree value were docked with AMPK and eNOS ，to verify the interaction. RESULTS Compared with before treatment ，the number and water content of feces were increased significantly in Mayuan tongbian zhitong decoction groups （P＜0.05）. Compared with blank group ，carbon powder propulsion rate of model group was decreased significantly （P＜0.05）； colonic structure was disordered ，and a large number of inflammatory cells were seen in submucosa ；the levels of NO and NOS in colon tissue as well as the protein expressions of AMPK ，eNOS，mTOR，Tsc-1，Tsc-2 and 4ebp were increased significantly （P＜0.05）. Compared with model group ，above indexes of Mayuan tongbian zhitong decoction groups （except for NOS in low-dose group ）were reserved significantly （P＜0.05）. In the molecular docking experiment ，the active components with the highest Degree values in C. sativa ，C. aurantium and R. glutinosa were（Z）-3-（4-hydroxy-3-methoxy-phenyl）-N-[2-（4-hydroxyphenyl）ethyl] acrylamide，nobiletin and stigmasterol. The binding energies of AMPK with these three components were －5.15，－4.61 and －4.83 kJ/mol，the binding energies of eNOS with these three components were －6.11，－5.40 and －5.91 kJ/mol. The conformations of these three compounds with AMPK and eNOS were stable and their binding activities were high. CONCLUSIONS Mayuan tongbian zhitong decoction can improve the constipation symptoms and intestinal function in STC model rats ，and its specific mechanism may be related to the inhibition of AMPK/eNOS signaling pathway.

Objective:To analyze the failure patterns and influencing factors of stereotactic ablative radiotherapy (SABR) for early-stage non-small cell lung cancer (ES-NSCLC).Methods:113 cases of ES-NSCLC treated with SABR from 2012 to 2020 in our hospital were retrospectively analyzed. The failure patterns, recurrence time, recurrence site and influencing factors were analyzed. Kaplan-Meier method was used to calculate the local recurrence rate, regional lymph node recurrence rate and distant metastasis rate. Univariate analysis was performed by Log-rank test, and multivariate analysis was performed by Cox model.Results:The median follow-up time was 58 months (range: 6-108 months), and a total of 45 patients (39.8%) recurred. The median recurrence time was 36 months. Distant metastasis (DM) occurred in 31 patients (27.4%) and DM alone in 24 patients (21.2%). Local recurrence (LR) was developed in 12 patients (10.6%) and LR alone in 7(6.2%). Regional lymph node recurrence (RR) occurred in 11 patients (9.7%) and RR alone in 6 patients (5.3%). LR combined with RR was observed in 1 case (0.9%), LR combined with DM in 3(2.7%), LR combined with RR and DM in 1(0.9%), and RR combined with DM in 3(2.7%). The 1-, 2-, 3-, 4-and 5-year recurrence rates were 5.4%, 16.6%, 27.5%, 44% and 51.2%, respectively. Univariate and multivariate analyses suggested that EGFR mutation was an influencing factor of high recurrence rate.Conclusion:ES-NSCLC patients treated with SABR alone have a high recurrence rate, and DM is the most common mode of failure. Follow-up consolidation therapy is recommended, especially for EGFR mutation-positive NSCLC patients.

Atherosclerosis (AS) is a chronic and progressive arterial disease. It is an important cause of the occurrence and development of cardiovascular and cerebrovascular diseases. With the development of Traditional Chinese Medicine (TCM), TCM has many advantages in the therapy of AS, with less adverse reactions. Studies have shown that TCM can resist AS, and the mechanism mainly belongs to regulating lipid metabolism, anti-lipid peroxidation, anti-inflammation, anticoagulation, and protecting the structure and function of vascular endothelial cells. The mechanism of TCM for AS is warranted to be studied systematically, and the chemical components need to be further clarified.

Objective:To analyze the clinical efficacy of different treatment modalities and prognostic factors of patients with Masaoka-Koga stage Ⅲ thymoma.Methods:Clinical data of patients diagnosed with Masaoka-Koga stage Ⅲ thymoma admitted to Affiliated Cancer Hospital of Zhengzhou University from January 2000 to December 2018 were analyzed retrospectively. A total of 133 patients had complete treatment and follow-up data. Kaplan-Meier method was used to calculate the cumulative survival rate, log-rank method was used to compare the survival between two groups, and Cox regression model was used for multivariate analysis.Results:The median follow-up time was 50 months (3-221 months). The median overall survival (OS) was 51 (3-221) months, and the median disease-free survival (DFS) was 45 (2-221) months. The survival rate in the radical surgery group was better than that in the palliative surgery group. The 5- and 10-year OS rates in radical surgery group were 88.2% and 74.4% respectively, while in palliative surgery group were 51.8% and 32.4% respectively ( P<0.001). The 5- and 10-year DFS rates in radical surgery group were 72.2% and 45.5%, respectively, while in palliative surgery group were 32.3% and 16.1% respectively ( P=0.001). The OS in the surgery combined with radiotherapy group was better than that in the surgery alone group. The 5- and 10-year OS rates in the radical surgery group were 82.8% and 64.2% respectively, while in the palliative surgery group were 55.8% and 50.2% ( P=0.033). There was no significant difference in DFS between two groups ( P=0.176). Multivariate analysis showed that age < 50 years old ( HR=0.264, P=0.001), radical resection ( HR=0.134, P<0.001), surgery combined with radiotherapy ( HR=2.778, P=0.009) were independently associated with better OS. Age < 50 years old ( HR=0.550, P=0.046), radical resection ( HR=0.555, P=0.042), and invasion of single organ ( HR=0.111, P=0.003) were independently associated with better DFS. Conclusions:OS and DFS in patients undergoing radical surgery are significantly better than those in their counterparts treated with palliative surgery, which is the most important factor affecting prognosis. Surgery combined with radiotherapy yields better OS. It is necessary to design a rigorous and reasonable multicenter prospective study to evaluate the efficacy of various treatment modalities and prognostic factors.

Objective:To analyze the invasion characteristics and prognostic factors of patients with Masaoka-Koga stage Ⅲ thymoma.Methods:The tumor invasion characteristics of 179 patients who were diagnosed with Masaoka-Koga stage Ⅲ thymoma and treated in Affiliated Cancer Hospital of Zhengzhou University from January 2000 to June 2018 were analyzed retrospectively. According to the treatment methods, all patients were divided into the radical operation group ( n=94), palliative operation group ( n=39) and simple biopsy group ( n=46). The χ2 test was used to compare the classified variables, Kaplan- Meier method was utilized to calculate the cumulative survival rate, log-rank method was used for group comparison and univariate analysis, and Cox’s regression model was used for multivariate analysis. Results:Mediastinal pleural invasion (86.0%) was the most common site, followed by pericardium (50.8%), great vessel (40.8%) and lung (36.3%). The proportion of macrovascular invasion in the radical operation group was 14.9%, significantly lower than 79.5% and 60.9% in the palliative surgery group and biopsy group (both P<0.001). Multivariate analysis showed that the nature of operation ( P<0.001), age ( P=0.011), radiotherapy ( P=0.020) were the independent factors affecting overall survival (OS), while nature of operation ( P<0.001), age ( P=0.004), radiotherapy ( P=0.020), number of invasive organs ( P=0.023) and pathological type ( P=0.016) were the independent factors affecting progress-free survival (PFS). Conclusions:For patients with Masaoka-Koga stage Ⅲ thymoma, mediastinal pleura is the most common site of invasion, pericardium, lung and great vessels are also commonly invaded. The invasion of mediastinal pleura, pericardium and lung exerts slight effect on surgical resectability, whereas great vessel involvement can significantly affect surgical resectability. OS and PFS in patients undergoing radical resection are significantly better than those in patients treated with palliative resection and biopsy. Radical resection is the most important factor affecting prognosis.

Objective:To evaluate the efficacy and safety of stereotactic ablative radiotherapy (SABR) in patients with pulmonary oligometastases.Methods:Clinical data of patients with pulmonary metastases treated with SABR from 2011 to 2018 were retrospectively analyzed. The local control rate (LCR) and overall survival (OS) were calculated by Kaplan- Meier method. log-rank test was used for univariate analysis and Cox’s regression model for multivariate analysis. Results:A total of 214 lung metastases were detected in 159 patients, and the median follow-up time was 43 months. The 1-, 3-and 5-year LCR were 90.1%, 73.9% and 65.8%, respectively. The 1-, 3-and 5-year OS were 73.8%, 43.6% and 11.9%, respectively. Univariate analysis showed that biological effective dose (BED)&ge;100 Gy was significantly correlated with LCR ( P=0.033). Cox’s multivariate analysis showed that BED and primary tumor source were the independent prognostic factors of LCR ( P=0.023, P=0.043). No>grade 3 adverse events were observed in all patients during treatment. Conclusions:SABR is a safe and effective treatment of lung oligometastases. SABR should be actively aD ministered for pulmonary oligometastases, especially for those with lesions from lung cancer and the radiation dose should be selected as BED &ge;100 Gy.