ObjectiveTo investigate the chemical constituents of Chuanxiong Rhizoma-Paeoniae Radix Rubra（CRPRR） that cross the blood-brain barrier in rats with ischemic stroke， their brain-protective effects， and their impact on inflammatory factors including tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18） based on ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS） and pharmacodynamic experiments. MethodsA focal cerebral ischemia-reperfusion injury model was established in rats via the middle cerebral artery occlusion/reperfusion （MCAO/R） method using intraluminal suture. Neurological function was evaluated using behavioral scoring. UPLC-Q-TOF-MS was employed to identify the chemical constituents of CRPRR that crossed the blood-brain barrier and entered the cerebrospinal fluid in MCAO/R model rats. Male Sprague-Dawley rats were randomly divided into six groups： sham operation group， model group， low-， medium-， and high-dose CRPRR groups （1.35， 2.7， 5.4 g·kg^-1， respectively）， and an edaravone group （5 mg·kg^-1）， with 12 rats in each group. The sham and model groups received normal saline， while the treatment groups received the respective doses of CRPRR once daily by gavage for three consecutive weeks. The brain-protective effects of CRPRR were assessed using the Longa five-point scoring method， open field test， Morris water maze， 2，3，5-triphenyltetrazolium chloride （TTC） staining， hematoxylin and eosin （HE） staining， and transmission electron microscopy. ResultsNine chemical constituents were identified in the cerebrospinal fluid containing CRPRR， namely paeoniflorin， senkyunolide F， senkyunolide G， paeonimetabolin Ⅰ， paeoniflorin derivative， senkyunolide H， benzoylpaeoniflorin， senkyunolide A， and ligustilide. Animal experiment results showed that compared with the sham operation group， the model group exhibited disordered neuronal arrangement， severe vacuolation， nuclear pyknosis， and evident mitochondrial swelling. Chromatin aggregation and peripheralization were also observed. Neurological scores and the number of crossings in the central region were significantly increased （P<0.01）， while platform crossings were significantly decreased （P<0.01）， and clear infarct areas were present （P<0.01）. Serum levels and protein expression of TNF-α， IL-1β， and IL-18 were significantly elevated （P<0.01）. Compared with the model group， all dose groups of CRPRR showed marked improvement in neuronal morphology which was close to the normal level， with mitochondrial swelling alleviated and chromatin distribution more uniform. The medium- and high-dose groups significantly reduced neurological scores （P<0.01）， while the low-， medium-， and high-dose groups significantly reduced the number of central crossings （P<0.01） and infarct volume （P<0.01）， and decreased TNF-α， IL-1β， and IL-18 levels （P<0.05， P<0.01） compared with the model group. Furthermore， the medium- and high-dose groups significantly reduced TNF-α protein expression （P<0.05，P<0.01）， and the high-dose group significantly reduced IL-1β and IL-18 protein expression （P<0.01）. ConclusionThis study confirmed that CRPRR improves neurological function and alleviates brain tissue damage in MCAO/R rats. Its mechanism may be associated with the downregulation of inflammatory factors TNF-α， IL-1β， and IL-18， as well as the presence of nine active chemical constituents in cerebrospinal fluid， namely paeoniflorin， senkyunolide F， senkyunolide G， paeonimetabolin Ⅰ， paeoniflorin derivative， senkyunolide H， benzoylpaeoniflorin， senkyunolide A， and ligustilide， which are closely related to their brain-protective effects.

ObjectiveTo investigate the chemical constituents of Chuanxiong Rhizoma-Paeoniae Radix Rubra（CRPRR） that cross the blood-brain barrier in rats with ischemic stroke， their brain-protective effects， and their impact on inflammatory factors including tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18） based on ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS） and pharmacodynamic experiments. MethodsA focal cerebral ischemia-reperfusion injury model was established in rats via the middle cerebral artery occlusion/reperfusion （MCAO/R） method using intraluminal suture. Neurological function was evaluated using behavioral scoring. UPLC-Q-TOF-MS was employed to identify the chemical constituents of CRPRR that crossed the blood-brain barrier and entered the cerebrospinal fluid in MCAO/R model rats. Male Sprague-Dawley rats were randomly divided into six groups： sham operation group， model group， low-， medium-， and high-dose CRPRR groups （1.35， 2.7， 5.4 g·kg^-1， respectively）， and an edaravone group （5 mg·kg^-1）， with 12 rats in each group. The sham and model groups received normal saline， while the treatment groups received the respective doses of CRPRR once daily by gavage for three consecutive weeks. The brain-protective effects of CRPRR were assessed using the Longa five-point scoring method， open field test， Morris water maze， 2，3，5-triphenyltetrazolium chloride （TTC） staining， hematoxylin and eosin （HE） staining， and transmission electron microscopy. ResultsNine chemical constituents were identified in the cerebrospinal fluid containing CRPRR， namely paeoniflorin， senkyunolide F， senkyunolide G， paeonimetabolin Ⅰ， paeoniflorin derivative， senkyunolide H， benzoylpaeoniflorin， senkyunolide A， and ligustilide. Animal experiment results showed that compared with the sham operation group， the model group exhibited disordered neuronal arrangement， severe vacuolation， nuclear pyknosis， and evident mitochondrial swelling. Chromatin aggregation and peripheralization were also observed. Neurological scores and the number of crossings in the central region were significantly increased （P<0.01）， while platform crossings were significantly decreased （P<0.01）， and clear infarct areas were present （P<0.01）. Serum levels and protein expression of TNF-α， IL-1β， and IL-18 were significantly elevated （P<0.01）. Compared with the model group， all dose groups of CRPRR showed marked improvement in neuronal morphology which was close to the normal level， with mitochondrial swelling alleviated and chromatin distribution more uniform. The medium- and high-dose groups significantly reduced neurological scores （P<0.01）， while the low-， medium-， and high-dose groups significantly reduced the number of central crossings （P<0.01） and infarct volume （P<0.01）， and decreased TNF-α， IL-1β， and IL-18 levels （P<0.05， P<0.01） compared with the model group. Furthermore， the medium- and high-dose groups significantly reduced TNF-α protein expression （P<0.05，P<0.01）， and the high-dose group significantly reduced IL-1β and IL-18 protein expression （P<0.01）. ConclusionThis study confirmed that CRPRR improves neurological function and alleviates brain tissue damage in MCAO/R rats. Its mechanism may be associated with the downregulation of inflammatory factors TNF-α， IL-1β， and IL-18， as well as the presence of nine active chemical constituents in cerebrospinal fluid， namely paeoniflorin， senkyunolide F， senkyunolide G， paeonimetabolin Ⅰ， paeoniflorin derivative， senkyunolide H， benzoylpaeoniflorin， senkyunolide A， and ligustilide， which are closely related to their brain-protective effects.

Objective:To develop a Core Competency Scale for Ophthalmic Specialist Nurses and test its reliability and validity.Methods:This study was a questionnaire development study. This study transformed the core competency evaluation indicators for ophthalmic specialist nurses constructed through literature review, semi-structured interviews, expert consultations, and other methods to form the initial draft of the Core Competency Scale for Ophthalmic Specialist Nurses. From October 2022 to March 2023, a survey was conducted on 364 ophthalmic specialist nurses in China using purposive and snowball sampling. Item analysis, exploratory factor analysis, and reliability testing were performed on the scale.Results:The final Core Competency Scale for Ophthalmic Specialist Nurses included six dimensions of ophthalmic specialized knowledge, ophthalmic specialized skills, essential nursing practice ability, communication and education ability, scientific research ability, and professional competence, totaling 30 items. Exploratory factor analysis extracted six common factors, with a cumulative variance contribution rate of 73.077%. The total Cronbach's α coefficient of the scale was 0.955, the half reliability coefficient was 0.796, and the retest reliability coefficient was 0.973.Conclusions:The Core Competency Scale for Ophthalmic Specialist Nurses has good reliability and validity, which can be used to evaluate the core competency of ophthalmic specialist nurses.

As a normal physiological substance,D-galactose can induce a process similar to natural brain aging in vivo and in vitro when administered excessively,and thus it is widely used to induce brain aging models in China and abroad.The model of brain failure induced by D-galactose has the advantages of a short modeling time,low cost,and significant effect.However,the induction mechanisms are complex and diverse,and the relationships between the mechanisms are unclear,which limit the practical applications of the model.This article reviews the in vivo metabolism of D-galactose and the various mechanisms involved in the induction of brain aging,as well as the links between the mechanisms,to provide a reference for the application and development of this model and the in-depth study of brain aging.

The PCR-amplified severe fever with thrombocytopenia syndrome virus(SFTSV)Gn-DⅢ-Ⅲ gene was inserted into the pET-30a(+)prokaryotic expression vector to generate the re-combinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ.The plasmid was transformed into E.coli BL21(DE3)for Gn-DⅢ-m protein expression and the expression conditions were optimized.The Gn-DⅢ-Ⅲ protein purified with Ni-NTA column affinity chromatography was applied as the captured antigen to establish an indirect ELISA method for the detection of SFTSV antibody.The results demonstrated that the recombinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ was successfully constructed as identified by PCR and sequencing.The recombinant protein SFTSV Gn-D m-Ⅲ was soluble ex-pression in E.coli under the optimal induction conditions of 0.4 mmol/L IPTG at 25 ℃ for 4 h,and the protein purity was 91.77％after purification by Ni-NTA column.The optimal reaction con-ditions for the indirect ELISA of SFTSV antibody were as follows:coating antigen concentration(5 μg/mL),primary antibody(incubation at 37 ℃ for 1.5 h),and secondary antibody(diluted 1:10 000 and incubated at 37 ℃ for 1 h).The established method had no cross-reactivity with Rift Valley fever virus(RVFV),Ebola virus(EBOV),and tick-borne encephalitis virus(TBEV)posi-tive sera.The method had a high sensitivity,with P/N＞2.1 for SFTSV-positive sera diluted to 81920.Coefficients of variation for intra-and inter-batch reactions were less than 10％.Detection of four SFTSV-infected human clinical serum samples showed the serum samples from patients in re-mission were tested as positive(P/N＞2.1),while serum samples from patients with multiple or-gan failure were detected as negative(P/N＜2.1).The results indicated that the SFTSV Gn-D Ⅲ-Ⅲ protein was successfully expressed and purified,and it was used as the coating protein to estab-lish an indirect ELISA assay for SFTSV antibody,which possesses good specificity,sensitivity and reproducibility.This method might be applied to detect human SFTSV clinical serum samples.

Objective:To explore the clinical features and prognosis of patients with primary central nervous system lymphoma(PCNSL).Methods:A retrospective analysis was performed on the relationship between clinical features,treatment regimen and prognosis in 46 newly diagnosed patients with primary central nervous system lymphoma who were diagnosed and treated in The Second Hospital of Lanzhou University from January 2015 to September 2022.Fisher's exact probability method was used to analyze the differences in clinical data of different subgroups.Kaplan-Meier survival curve was used to analyze the overall survival rate and progression-free survival rate of patients with different treatments,and the factors influencing survival were analyzed.Results:Among 46 patients with PCNSL,which pathological type were diffuse large B-cell lymphoma(DLBCL).There were 26(56.5％)cases of male and 20(43.5％)of female,with a median age of 54(17-71)years.In Hans subtypes,14 cases(30.4％)of GCB subtype,32 cases(69.6％)of non-GCB subtype.32 cases(69.6％)of Ki-67 ≥80％.Among 36 patients who completed at least 2 cycles of treatment with follow-up data,the efficacy evaluation was as follows:overall response rate(ORR)was 63.9％,complete response(CR)rate was 47.2％,17 cases of CR,6 cases of PR.The 1-year progression-free survival rate and 1-year overall survival rate was 73.6％and 84.9％,respectively.The 2-year progression-free survival rate and 2-year overall survival rate was 52.2％and 68.9％,respectively.The ORR and CR rate of 17 patients treated with RMT regimen was 76.5％and 52.9％(9 cases CR and 4 cases PR),respectively.Univariate analysis of 3 groups of patients treated with RMT regimen,RM-BTKi regimen,and RM-TT regimen as first-line treament showed that deep brain infiltration was associated with adverse PFS(P=0.032),and treatment regimen was associated with adverse OS in PCNSL patients(P=0.025).Conclusion:Different treatment modalities were independent prognosis predictors for OS,the deep brain infiltration of PCNSL is a poor predictive factor for PFS.Patients with relapse/refractory(R/R)PCNSL have a longer overall survival time because to the novel medication BTKi.They have strong toleration and therapeutic potential as a first-line therapy for high-risk patients.

Objective:To investigate the clinical characteristics and influence of co-mutated gene on acute myeloid leukemia patients(AML)with FMS-like tyrosine kinase-3(FLT3)mutations.Methods:A total of 273 FLT3+AML patients were enrolled,and the co-mutation gene data of the patients were collected to further analyze the prognosis of the patients.FLT3 and other common mutations were quantified by PCR amplification products direct sequencing and second-generation sequencing(NGS).Results:When patients were divided into FLT3 ITD+,FLT3 TKD+,FLT3 ITD++TKD+and FLT3 ITD-+TKD-group according to the type of FLT3 mutations,it was found that the frequencies of TET2,GATA2,NRAS and ASXL1 mutation were significantly different among the 4 groups(all P＜0.05).When patients were divided into allelic ratio(AR)≥0.5 and＜0.5 group,it was found that the frequencies of FLT3 ITD+,FLT3 ITD-+TKD-,NPM1,NRAS and C-kit were significantly different between the two groups(all P＜0.05).When patients were divided into normal and abnormal karyotype group,it was found that the frequencies of FLT3 ITD+,FLT3 TKD+,NPM1,GATA2 and C-kit were significantly different between the two groups(all P＜0.05).The median overall survival(OS)of AML patients with FLT3 TKD+(including FLT3 ITD++TKD+)was longer than that of patients with FLT3 ITD+alone(P＜0.05).The OS and relapse-free survival(RFS)of AML patients with FLT3++TET2+were both shorter than those of patients with FLT3++TET2-(both P＜0.05).Conclusion:The mutation frequencies of co-mutated genes are correlated with subtypes of FLT3,karyotype and AR.AML patients with FLT3 TKD+have longer OS than patients with FLT3 ITD+alone,and patients with co-mutation of TET2 have shorter median OS and RFS.

Objective:To explore the effect of heterozygous deletion of histone methyltransferase Kmt2c gene on the hematological system of mice.Methods:CRISPR/Cas9 technology was used to construct mice model of Kmt2c heterozygous deletion(Kmt2c+/-)and the changes of whole blood cell count in mice were continuously monitored by blood routine test.The clonal expansion ability of bone marrow cells was explored by colony formation assay in vitro and the proportion of primitive hematopoietic cells,including long-term hematopoietic stem cell(LT-HSC),short-term hematopoietic stem cell(ST-HSC),and multipotent progenitor cell in mutant mice was analyzed by flow cytometry.Results:Kmt2c+/-mice model was successfully constructed,and the mRNA expression level of Kmt2c was 28％of that of C57BL/6J mice.The colony formation ability of bone marrow cells of Kmt2c+/-mice in vitro increased with the passage times,and the colony number in the fourth generation was significantly higher than that of control group(P＜0.05).The proportions of LT-HSC and ST-HSC in the primitive hematopoietic cell population of Kmt2c+/-mice was 19.6％±3.3％and 28.9％±4.9％,respectively,which showed an increasing trend compared with 16.9％±2.6％and 18.9％±2.5％in control group,but the difference was not statistically significant(P＞0.05).The white blood cell count of Kmt2c+/-mice gradually increased after 12 weeks of monitoring and reached(9.8±1.0)×109/L at the 14th week,which was significantly higher than(7.3±1.4)× 109/L of control group(P＜0.05).Conclusion:The bone marrow cells of Kmt2c+/-mice have potential of clonal expansion.

Objective:To determine the serum levels of CMTM6 mRNA and CCN1 in patients with acute leukemia (AL),and to analyze their relationship with the clinical efficacy and prognosis of the patients. Methods:103 AL patients admitted to our hospital from February 2015 to January 2019 were included as the study subjects. Additionally,100 healthy subjects who underwent physical examinations during the same period were included as the control group. qRT-PCR method was applied to detect the serum CMTM6 mRNA level of the study subjects,the serum CCN1 level was measured by ELISA. The levels of serum CMTM6 mRNA and CCN1 between the control group and AL patients,as well as between patients at initial diagnosis and after one course of chemotherapy were compared,the correlation of CMTM6 mRNA and CCN1 levels at initial diagnosis with clinicopathological features and short-term efficacy in AL patients was analyzed. The correlation of the CMTM6 mRNA and CCN1 expression levels with prognosis of the patients was analyzed by Kaplan-Meier curves. Results:Compared with the control group,the serum CMTM6 mRNA level in AL patients was significantly increased (P<0.05),while the serum CCN1 level was significantly decreased (P<0.05). There were no statistically significant differences in serum CMTM6 mRNA and CCN1 levels between patients with acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML) (P>0.05). Compared with those at the initial diagnosis,both the serum CMTM6 mRNA and CNN1 expression levels in AL patients were significantly altered after one course of chemotherapy,with CMTM6 mRNA significantly decreased (P<0.05) and CCN1 significantly increased (P<0.05). The 3-year recurrence-free survival (RFS) rate was lower in the CMTM6 high expression group and the CCN1 low expression group,compared with the CMTM6 low expression group and CCN1 high expression group,respectively (P<0.05). Conclusion:Serum CMTM6 is highly expressed and CCN1 is lowly expressed in AL patients. The expression levels of serum CMTM6 and CCN1 are associated with the 3-year prognosis of the patients.

Objective:To investigate the clinical efficacy of antiperistaltic uncut Roux-en-Y anastomosis (Uncut RY) in laparoscopic distal gastrectomy (LDG) for gastric cancer (GC).Methods:The clinicpathological data of 309 patients who underwent LDG for GC between Jan 2020 to Dec 2021 in Tianjin Medical University Cancer Institute & Hospital were collected. Among them, 102 cases received antiperistaltic Uncut RY anastomosis, 96 cases did RY anastomosis, and 111 cases did B Ⅱ+Braun anastomosis. The general data and clinical efficacy were compared.Results:The operative time in Uncut RY group and B Ⅱ+Braun group was shorter than that in RY group ( F=3.356, P=0.036). The incidence of postoperative short-term complications was not statistically different among three groups ( P>0.05). Compared to Uncut RY group and RY group, the B Ⅱ+Braun group had a significantly higher incidences of reflux gastritis (19.8% vs. 6.9% vs.5.2%, F=13.889, P=0.001). The incidence of RY stasis syndrome in RY group (15.6%) was significantly higher than that in Uncut RY group (2.0%) and B Ⅱ+Braun group (2.7%) ( F=19.320 ,P=0.000). There were no significant differences in postoperative 1-year nutritional indexes among three group ( P>0.05). Conclusions:The antiperistaltic Uncut RY anastomosis in LDG for GC is safe and feasible, which can reduce the incidences of reflux gastritis and RY stasis syndrome.