ObjectiveTo establish fingerprint profiles and a quantitative determination method for Lycii Cortex， providing a scientific basis for the formulation of quality standards for Lycii Cortex and its roasted products. MethodsHigh performance liquid chromatography（HPLC） was developed for the quantitative method for determining kukoamine B in Lycii Cortex and its roasted products on an Alphasil XD-C₁₈ CH column（4.6 mm×250 mm， 5 μm）. HPLC fingerprint profiles were established for 10 batches of Lycii Cortex and its roasted products， and ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was used to identify the common peaks based on reference standards， literature and MS information. Quality evaluation indicators included yield of decoction pieces， appearance properties， content of kukoamine B， and fingerprint profiles. The temperature and time of the roasting process were investigated to select the optimal preparation process， which was then verified. Additionally， chemical pattern recognition was combined to assess the differences in the chemical composition of Lycii Cortex before and after roasting， as well as among samples from different origins. ResultsQuantitative analysis indicated that the contents of kukoamine B in Lycii Cortex and its roasted products were 0.35%-5.51% and 0.24%-4.15%， respectively. The transfer rate of kukoamine B was 58.6%-78.9% after roasting. The fingerprint profile analysis demonstrated that the method established in this study effectively separated kukoamine B from other components in the samples and distinctly differentiated it from its impurity peak， cis-N-caffeoylputrescine. The HPLC fingerprint profiles of Lycii Cortex and its roasted products showed high similarity（all above 0.95）， with 7 common peaks identified and five common components， including kukoamine B， cis-N-caffeoylputrescine， N-coumaroyl tyramine， feruloyltyramine， and glucosyringic acid， confirmed. Process optimization confirmed that baking at 110 ℃ for 20 min was a stable and feasible method for roasting Lycii Cortex. Principal component analysis and cluster analysis showed that there was little difference in the chemical composition between raw and roasted Lycii Cortex， but the quality of Lycii Cortex from different origins differed greatly. ConclusionThis study successfully established the fingerprint profiles and a quantitative method for the effective component kukoamine B in Lycii Cortex and roasted Lycii Cortex. The qualitative and quantitative analyses clarified that the impact of the roasting process on the chemical composition of Lycii Cortex was less significant than the variations due to its geographical origin. The findings of this study offer a reference for the development of quality evaluation methods and the establishment of quality standards for Lycii Cortex and its processed products.

Multiple myeloma (MM) is a hematological malignancy that poses significant treatment challenges due to its heterogeneity and propensity for relapse and progression. In the last two decades, the therapeutic landscape of MM has changed dramatically, but the disease remains largely incurable, with many patients facing treatment resistance. This review evaluates the current status of MM treatments, emphasizing the limitations of traditional therapies and the emerging role of immunotherapy in improving patient outcomes. It highlights the importance of achieving and maintaining minimal residual disease negativity and a balanced immune response as key treatment goals. Furthermore, it discusses the advancements in immunotherapies that are improving the prospects for patients, particularly those with relapsed or refractory disease. Innovative strategies, such as chimeric antigen receptor T-cell therapy, bispecific antibodies, and bispecific T cell engagers, have shown significant promise by targeting the malignant cells and the bone marrow microenvironment, which are essential for disease persistence and resistance to therapy. Future research should focus on refining MM treatment strategies, including the integration of immunotherapy into earlier treatment lines and the development of predictive biomarkers for personalized treatment approaches, ultimately enhancing patient outcomes.

Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering, which limit sequence and functional diversity, thereby constraining their therapeutic and application potential. Protein design tools have significantly advanced the creation of novel protein sequences, structures, and functions. However, further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability. Here, we extended an evolution-based design protocol to create a novel minibinder, BindHer, against the human epidermal growth factor receptor 2 (HER2). It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity. Radiolabeling experiments with ^99mTc, ⁶⁸Ga, and ¹⁸F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models, with minimal nonspecific liver absorption, outperforming scaffolds designed through traditional engineering. These findings highlight a new rational approach to automated protein design, offering significant potential for large-scale applications in therapeutic mini-protein development.

Objective To investigate the correlation between MEX3A and differentiation characteristics of gastric cancer and intestinal metaplasia,and its combination with caudal-related homeobox transcription factor 2(CDX2)and mucin 2(MUC2)and mucin 5AC(MUC5AC)to determine the role of carcinogenic intestinal metaplasia.Methods From January 2010 to December 2014,a total of 410 cases of gastric cancer and paracarcinoma paraffin-embedded tissue samples were selected from the Central Hospital Affiliated to Shenyang Medical College and the Second Hospital Affiliated to Shenyang Medical College.According to pathological diagnosis,they were divided into control group(mild superficial gastritis,79 cases),intestinal metaplasia group(149 cases)and gastric cancer group(182 cases).The expressions of MEX3A,CDX2,MUC2 and MUC5AC were detected by immunohistochemistry.Results MEX3A was highly expressed in gastric cancer group and intestinal metaplasia group,especially diffuse gastric cancer,poorly differentiated gastric cancer and type Ⅲ intestinal metaplasia(P<0.05).CDX2 and MUC2 were highly expressed in gastric cancer group and intestinal metaplasia group,especially intestinal type gastric cancer,highly and moderately differentiated gastric cancer,type Ⅰ and type Ⅱ intestinal metaplasia(P<0.05).The expression of MUC5AC was high in control group and low in gastric cancer group and intestinal metaplasia group,especially in intestinal type gastric cancer,type Ⅰ and type Ⅲ intestinal metaplasia(P<0.05).Gastric cancer and intestinal metaplasia differentiation were negatively correlated with MEX3A and MUC5AC expression,but positively correlated with CDX2 and MUC2 expression(P<0.05).MEX3A was negatively correlated with the expression of CDX2 and MUC2,and positively correlated with the expression of MUC5AC in gastric cancer(P<0.05).MEX3A was negatively correlated with the expression of CDX2 and MUC2 in intestinal metaplasia(P<0.05),while CDX2 was positively correlated with the expression of MUC2(P<0.05).Conclusion MEX3A is negatively correlated with gastric cancer and intestinal metaplasia differentiation.Gastric cancer is characterized by high MEX3A expression and low CDX2 and MUC2 expression.

Objective To investigate the dietary patterns of rural residents in the high-incidence areas of esophageal cancer (EC), and to explore the clustering and influencing factors of risk factors associated with high-incidence characteristics. Methods A special structured questionnaire was applied to conduct a face-to-face survey on the dietary patterns of rural residents in Yanting county of Sichuan Province from July to August 2021. Univariate and multivariate logistic regression models were used to analyze the influencing factors of risk factor clustering for EC. Results There were 838 valid questionnaires in this study. A total of 90.8% of rural residents used clean water such as tap water. In the past one year, the people who ate fruits and vegetables, soybean products, onions and garlic in high frequency accounted for 69.5%, 32.8% and 74.5%, respectively; the people who ate kimchi, pickled vegetables, sauerkraut, barbecue, hot food and mildew food in low frequency accounted for 59.2%, 79.6%, 68.2%, 90.3%, 80.9% and 90.3%, respectively. The clustering of risk factors for EC was found in 73.3% of residents, and the aggregation of two risk factors was the most common mode (28.2%), among which tumor history and preserved food was the main clustering pattern (4.6%). The logistic regression model revealed that the gender, age, marital status and occupation were independent influencing factors for the risk factors clustering of EC (P<0.05). Conclusion A majority of rural residents in high-incidence areas of EC in Yanting county have good eating habits, but the clustering of some risk factors is still at a high level. Gender, age, marital status, and occupation are influencing factors of the risk factors clustering of EC.

Porcine deltacoronavirus (PDCoV) is a major pathogen causing fatal diarrhea in suckling piglets, and there is currently a lack of effective vaccines and drugs to prevent and control the virus. The nonstructural protein 13 (NSP13) serves as a virus-coded helicase and is considered to be a crucial target for antiviral drugs, making it imperative to investigate the helicase activity of NSP13. In this study, the NSP13 gene of PDCoV was synthesized and integrated into the prokaryotic expression vector pET-28a to construct the recombinant plasmid pET-28a-NSP13. NSP13 was successfully expressed in BL21 (DE3) and subsequently purified. The study also verified the helicase activity of the purified NSP13 and explored the factors that influence this activity. The results indicated that NSP13 from PDCoV was effectively expressed in the prokaryotic system and exhibited helicase activity, capable of unwinding double-stranded DNA with a tail at the 5' end. Additionally, NSP13 demonstrated an annealing function by promoting the complementary pairing of single-stranded nucleotide chains to form double strands. The helicase activity of NSP13 was affected by metal ions, but Mg²⁺concentrations in the range of 0.5-6.0 mmol/L had no significant effect on helicase activity of NSP13. When the solution pH was in the range of 4-9, there was no difference in helicase activity. ATP concentrations in the range of 0.25-6.00 mmol/L had a weak effect on helicase activity, and NSP13 concentration ≥80 nmol/L inhibited the helicase activity. We obtained the NSP13 of PDCoV and investigated its helicase activity. These findings provided a theoretical foundation for the further research on the regulatory mechanism of NSP13 in PDCoV replication and the development of anti-coronaviral drugs.
Viral Nonstructural Proteins/metabolism* ; Escherichia coli/metabolism* ; Recombinant Proteins/metabolism* ; Swine ; Animals ; DNA Helicases/metabolism* ; Genetic Vectors/metabolism*

AIM: To evaluate the dose regimens of tegacycline for treatment of hospital-acquired pneumonia, complex abdominal infection and complex skin and soft tissue infection caused by Gram-negative bacterial infections with Monte Carlo model. METHODS: The minimum inhibitory concentrations (MICs) of tegacycline against Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae and Enterobacter cloacae were collected from the CHINET in 2018. The target probability (PTA) and cumulative response fraction (CFR) of different regimens were calculated using Monte Carlo simulation based on PK/PD theory of tegacycline. RESULTS:In the treatments of HAP caused by gram-negative bacteria, when MIC≤0.5 μg / mL, the PTA of 50 mg q12h was greater than 90%, and when MIC≥1 μg / mL, PTA and CFR of 100 mg q12h were both greater than 90%, When MIC≥2 μg/mL, 50 mg q12h, 75 mg q12h and 100 mg q12h doses of PTA were less than 90%. In the treatment of cIAI, when MIC≤0.5 μg / mL, PTA of 50 mg q12h reached the target value, and when MIC=1 μg/mL, PTA of 100 mg q12h was greater than 90%. For complex skin and soft tissue infection, when the MIC≤0.25 μg/mL, the PTA of 75 mg q12h and 100 mg q12h was greater than 90%, and the PTA of the three administration regimen was less than 90%, when the MIC≥0.5 μg/mL. CONCLUSION:The dose of 50 mg q12h is more suitable for the treatment of HAP, when MIC>0.5 μg/mL, tigecycline may need 100 mg q12h to obtain the best clinical efficacy in the treatment of cIAI. For cSSSI. when MIC≤0.25 μg/mL, tigecycline can be administered with 75 mg q12h and 100 mg q12h. For the three types of infections caused by Escherichia coli, the conventional dose of tigecycline may achieve clinical efficacy.

Objective:To evaluate the efficacy and safety of autologous hematopoietic stem cell transplantation(auto-HSCT)for the treatment of multiple myeloma(MM)in elderly patients aged over 65 years.Methods:In this retrospective analysis, the efficacy and safety of auto-HSCT for the treatment of MM was examined in 28 patients aged >65 years diagnosed and treated at Soochow Hopes Hematology Hospital between March 1, 2020 and October 31, 2022.The functions of the major organs of these patients were evaluated before transplantation.Results:The 28 patients had a median age of 67(66-72)at the time of transplantation, a median number of 2.985 × 10 6/kg(2.036-9.5 × 10 6/kg)of collected CD34+ hematopoietic stem cells, and a median number of 2(1-3)days of collection.The median time to neutrophil implantation after hematopoietic stem cell transfusion was 10(9-14)days, and the median platelet implantation time was 11(10-29)days.The median follow-up time was 25 months, but the median progression-free survival time was not reached.The 1-year PFS rate was 89.3% and 2-year PFS rate was 76.3%, with 2 patients' starting point of PFS set at the time of pre-transplantation re-induction therapy because needed salvage auto-HSCT.The overall survival time was not reached, the 1-year overall survival rate was 100.0%, and the 2-year overall survival rate was 90.5%. Conclusions:Auto-HSCT is a safe and effective treatment for elderly MM patients aged over 65 years after screening and assessment.

Objective:To examine the long-term survival outcomes and identify mortality risk factors for coronary artery bypass grafting patients who received venoarterial extracorporeal membrane oxygenation(VA-ECMO) for postcardiotomy cardiogenic shock.Methods:Data from 121 consecutive venoarterial extracorporeal membrane oxygenation-treated coronary artery bypass grafting(CABG) patients at Beijing Anzhen Hospital between June 2012 and December 2016 were analyzed. There were 84 males and 24 females. The median age was 62(55, 67) years, and the median body mass index was 25(23, 27)kg/m 2. Thirty-seven patients(34%) had left main artery disease, and 56 patients(52%) underwent off-pump CABG. According to the 3 year survival outcome after ECMO, patients were divided into survival group(35 cases) and non-survival group(73 cases). The differences of clinical indicators between the two groups were compared and analyzed. Multivariable Cox regression modeling was used to identify factors independently associated with 36 month mortality. Results:Sixty-five patients(60%) could be weaned from VA-ECMO, 49 patients(45%) survived to hospital discharge, and 35 patients(32%) survived to 3 years. The median( IQR) time on VA-ECMO support was 4(3, 5) days. The median( IQR) length of ICU stay and hospital stay duration were 8(5, 12) and 20(13, 29) days, respectively. Older age( HR=1.06, 95% CI: 1.03-1.10, P<0.001), left main coronary artery disease( HR=1.62, 95% CI: 1.00-2.60, P=0.048), and vasoactive inotropic score( HR=1.09, 95% CI: 1.03-1.17, P=0.007)were independent risk factors associated with 3-year mortality. The area under the receiver operating characteristic curve for the model, which was constructed with age≥60years, left main coronary artery disease, and vasoactive inotropic score>60, was 0.88(95% CI: 0.80-0.95). Conclusion:Long-term survival of patients who survive to discharge appears favorable. Older age, left main coronary artery disease, and vasoactive inotropic score were associated with 3-year mortality in coronary artery bypass grafting patients who received VA-ECMO.

Objective:This study aimed to evaluate the efficacy and safety of lenalidomide combined with bortezomib and dexamethasone (VRD) in the treatment of newly diagnosed multiple myeloma (MM) .Methods:A total of 150 newly diagnosed patients with MM diagnosed in The First Affiliated Hospital of Soochow University from November 2018 to February 2021 and received VRD as the induction regimen were included to evaluate the safety and efficacy of VRD induction therapy for newly diagnosed MM.Results:The median follow-up was 22 months, two patients (1.3%) died early after treatment, and 148 patients (98.7%) completed induction therapy. 116 patients (77.3%) were mobilized to collect autologous hematopoietic stem cells, 101 cases (87.1%) were qualified in the collection, of which 48 cases (41.4%) were excellent in the collection. The 3-year progression-free survival (PFS) rate was 59%, and the 3-year overall survival (OS) rate was 83%. After induction, complete remission (CR) /stringent CR rate was 54.4%, ≥ very good partial remission rate was 77.3%, overall response rate was 86.0%, and minimal residual disease negative rate was 46.0%. There was no statistically significant difference in the efficacy of cytogenetic high-risk patients compared with standard risk patients ( P=0.456) . The median PFS time of cytogenetic high-risk patients was shorter than that of standard risk patients (not reached vs 33 months, P=0.014) . There was no statistically significant difference in the median OS time (not reached vs not reached, P=0.072) . The highest incidence of hematological adverse events was thrombocytopenia (72%) , followed by neutropenia (42%) and anemia (20%) . The highest incidence of non-hematological adverse events was peripheral neuritis (56.7%) . The main digestive tract symptoms include constipation (30.0%) and diarrhea (17.3%) . Upper respiratory tract infection (23.3%) and lung infection (7.3%) are the main infections. The incidence of adverse thrombocytopenia (90.0% vs 63.7%, P=0.001) , neutropenia (54.2% vs 36.3%, P=0.038) , anemia (33.3% vs 13.7%, P=0.005) , diarrhea (27.1% vs 12.7%, P=0.030) , limb edema (20.8% vs 3.9%, P=0.030) , fever (20.8% vs 4.9%, P=0.006) , thrombosis (8.3% vs 0, P=0.016) , and renal function deterioration (20.8% vs 3.9%, P=0.030) in patients with renal insufficiency was higher than that in patients with normal renal function. Conclusion:The VRD regimen has a significant effect on newly diagnosed MM, does not affect the hematopoietic stem cell collection, and has controllable adverse events; however, the incidence of adverse events was higher in patients with renal insufficiency.