[Objective] To establish a method for detecting the potency of recombinant human coagulation factor Ⅶa for injection. [Methods] By adding the sample and factor Ⅶ deficient plasma to the sample cup and activating the reaction with prothrombin time assay reagent (PT reagent), the coagulation time of the sample was determined by the change in magnetic bead swing amplitude in the sample cup. The logarithm of coagulation time was inversely proportional to the logarithm of human factor Ⅶa potency. [Results] Under the experimental conditions, the specificity of the methodology was evaluated through spiked recovery, and the recovery rates ranged from 90.0% to 110.0%. Within the range from 0.125 to 1.000 IU/mL, there was a good linear response between the potency and coagulation time of the standard and sample, with correlation coefficients r>0.99. As for the accuracy and repeatability, the recovery rates of various concentrations detected in the stock solution were 101.0%, 100.0% and 112.0%, respectively, with RSD values of 2.6%, 4.0% and 0.0%, respectively. The recovery rates of various concentrations in finished product testing were 104.0%, 94.7% and 112.0%, respectively, with RSD values of 1.9%, 2.4% and 0.0%, respectively. As for the intermediate precision, the RSD were 4.5% and 3.7%, respectively. After treated with sample diluent, the sample was tested at room temperature for 6 hours and still exhibited relatively stable biological activity. [Conclusion] This detection method is accurate, stable, easy to operate and highly automated, and is suitable for detecting the potency of recombinant human coagulation factor Ⅶa for Injection.

OBJECTIVE To observe the short-term efficacy and safety of venetoclax combined with homoharringtonine and cytarabine in the treatment of acute myeloid leukemia （AML）. METHODS The data of 40 newly diagnosed AML patients admitted to our hospital from October 2022 to November 2023 were retrospectively collected and divided into observation group and control group according to treatment plan， with 20 cases in each group. The patients in the control group were given Daunorubicin hydrochloride for injection+Cytarabine for injection， and the patients in the observation group were given Venetoclax tablets+ Homoharringtonine injection+Cytarabine for injection. The patients in both groups were given relevant medicine， with 28 days as one cycle. The short-term efficacy， negative rate of minimal residual disease （MRD）， duration of granulocyte deficiency， duration of platelet （PLT） ＜20×109 L－1， transfusion volume of suspended red blood cells and platelet， and the occurrence of adverse drug reactions were evaluated in both groups after 1 cycle of induction chemotherapy. RESULTS The complete remission or complete remission with incomplete hematologic recovery （CR/CRi） rate in the observation group was significantly higher than control group （P＜0.05）， and the negative rate of MRD in the observation group was also significantly higher than control group （P＜0.05）. However， in low-， medium- and high-risk patients， there was no statistical significance in CR/CRi rates between the two groups （P＞0.05）. There were no significant differences in the duration of agranulocytosis， the duration of PLT ＜20×109 L－1， the amount of suspended red blood cell transfusion， the amount of platelet transfusion， the incidence of hematologic toxicity and the incidence of non-hematologic toxicity between 2 groups （P＞0.05）. CONCLUSIONS Venetoclax combined with homoharringtonine and cytarabine show good short-term efficacy and safety in the treatment of AML.

Objective To evaluate the effects of transcutaneous electrical acupoint stimulation(TEAS)combined with Chinese medicine emotional intervention on immune function and psychological stress in patients undergoing laparoscopic radical rectal cancer surgery.Methods A total of 100 patients hospitalized for laparoscopic radical surgery for rectal cancer were randomly divided into 4 groups of Group A,B,C,and D,25 patients in each group.Group A was given TEAS for 30 minutes before anesthesia,Group B was given TEAS continuously until the end of the surgery,Group C was given TEAS for 30 minutes and Chinese medicine emotional intervention before anesthesia,Group D was given TEAS continuously until the end of the surgery together with Chinese medicine emotional intervention.TEAS was performed at bilateral Zusanli(ST36)points,bilateral Sanyinjiao(SP6)points,Baihui(GV20)points,bilateral Neiguan(PC6)points.The changes of perioperative outcomes and the changes of parameters associated with the immune function,molecular mechanism of Th1/Th2 cells,and psychological stress response before the surgery and 3 days after the surgery in each group were observed.Moreover,the occurrence of adverse reactions in each group was compared.Results(1)The differences of perioperative outcomes operation time,bleeding volume and infusion amount were not statistically significant among the 4 groups(P＞0.05).The Sufentanil dosage,Remifentanil dosage,postoperative awakening time,and 24-hour postoperative pain Visual Analogue Scale(VAS)scores of group D were lower than those of Groups A,B,and C(P＜0.05),and the above indexes of group B and group C were all lower than those of Group A(P＜0.05).(2)Three days after the surgery,the levels of immune function indicators of T lymphocyte subsets CD3+,CD4+ and natural killer(NK)cells in Group D were higher than those in Groups A,B,and C(P＜0.05),while the above indexes in Groups B and C were higher than those in Group A(P＜0.05).(3)Three days after the surgery,the levels of parameters associated with the molecular mechanism of Th1/Th2 cells such as interleukin 2(IL-2)and interferon gamma(IFN-γ)in Group D were higher(P＜0.05),and the serum interleukin 10(IL-10)level was lower than those in Groups A,B,and C(P＜0.05);the serum IL-2 and IFN-γ levels in Group B and C were higher(P＜0.05)and serum IL-10 level was lower than those in Group A(P＜0.05).(4)Three days after the surgery,the scores of the psychological stress response indicators of self-rating anxiety scale(SAS)and self-rating depression scale(SDS)in Group D were lower than those in Groups A,B,and C(P＜0.05),while the above scores in Groups B and C were lower than those in Group A(P＜0.05).(5)In Group A,there was one case of nausea and vomiting;in Group B,there were two cases of nausea and vomiting and one case of dizziness;no adverse reaction occurred in Group C,and there was one case of nausea and vomiting in Group D.The incidences of adverse reactions in Groups A,B,C and D were 4.0%(1/25),12.0%(3/25),0.0%(0/25),and 4.0%(1/25),respectively.The intergroup comparison showed no statistically significant difference among the 4 groups(χ2 = 0.400,P = 0.527).Conclusion In treating patients undergoing laparoscopic radical surgery for rectal cancer,TEAS combined with Chinese medicine emotional intervention can effectively regulate the perioperative immune function,reduce the stress response,and alleviate the anxiety of the patients.Moreover,the therapy exerts certain analgesic effect,which leads to the reduction of the dosage of opioids and the increase of perioperative comfort and surgical tolerance of the patients.

ObjectiveTo investigate the inhibitory effect of Mahuang Xixin Fuzitang on the migration of dendritic cells （DCs） in mice and its underlying mechanism. MethodMouse bone marrow-derived DCs were isolated and cultured. The morphological changes of the cells at different stages were observed under a microscope， and the CD11c⁺ proportion was detected by flow cytometry to identify DC purity. Cells were treated with Mahuang Xixin Fuzitang （1， 2， 5， 10， 20， 40， 50， 100 g·L^-1） for 24 hours， and the effect of Mahuang Xixin Fuzitang on cell proliferation was assessed using the cell counting kit-8 （CCK-8） assay to determine the appropriate concentrations for treatment. After modeling by lipopolysaccharide （LPS） induction， DCs were divided into a blank group， a model group， and Mahuang Xixin Fuzitang groups （2， 4， 8 g·L^-1）. The expression of surface molecules CD80， CD86， and major histocompatibility complex-Ⅱ （MHC-Ⅱ） were detected by flow cytometry. Transwell chamber assay was used to observe cell migration. The levels of chemokine C-C-primitive receptor 7 （CCR7） and chemokine C-X-C-primitive receptor 4 （CXCR4） on the cell surface were detected by enzyme-linked immunosorbent assay （ELISA）. The expression of filamentous actin （F-actin） in the cell microfilament cytoskeleton was detected by immunofluorescence （IF） staining. Real-time quantitative polymerase chain reaction （Real-time PCR） was employed to determine the mRNA expression levels of Ras homolog family member A （RhoA） and Rho-associated coiled-coil containing protein kinase 1 （ROCK1）. Western blot analysis was performed to detect the protein expression of RhoA and ROCK1. ResultCompared with the blank group， the model group exhibited significantly higher expression levels of CD80， CD86， and MHC-Ⅱ （P<0.01）， a significantly increased number of cells migrating to the lower chamber （P<0.01）， and significantly elevated levels of CCR7 and CXCR4 （P<0.05， P<0.01）. Additionally， F-actin expression was significantly increased （P<0.01）， and both RhoA and ROCK1 mRNA and protein expression levels were significantly higher （P<0.05， P<0.01）. Compared with the model group， treatment with Mahuang Xixin Fuzitang （2， 4， 8 g·L^-1） for 24 hours resulted in significantly lower expression levels of CD80， CD86， and MHC-Ⅱ （P<0.01）， a significantly reduced number of cells migrating to the lower chamber （P<0.05）， and significantly decreased levels of CCR7 and CXCR4 （P<0.05， P<0.01）. Furthermore， F-actin expression was significantly reduced （P<0.01）， and both RhoA and ROCK1 mRNA and protein expression levels were significantly decreased （P<0.05， P<0.01）. ConclusionMahuang Xixin Fuzitang can inhibit the migration of DCs in mice， and its mechanism of action may be related to reducing the activity of the Rho/ROCK signaling pathway， thereby affecting changes in the cell cytoskeleton.

Aim To investigate the effect of ellagic acid (EA) on cognitive function in APP/PS 1 double- transgenic mice, and to explore the regulatory mechanism of ellagic acid on the level of oxidative stress in the hippocampus of double-transgenic mice based on the phosphatidylinositol 3-kinase/protein kinase B/glycogen synthase kinase-3 (PI3K/AKT/GSK-3 β) signaling pathway. Methods Thirty-two SPF-grade 6-month-old APP/PS 1 double transgenic mice were randomly divided into four groups, namely, APP/PS 1 group, APP/PS1 + EA group, APP/PS1 + LY294002 group, APP/PS 1 + EA + LY294002 group, with eight mice in each group, and eight SPF-grade C57BL/6J wild type mice ( Wild type) were selected as the blank control group. The APP/PS 1 + EA group was given 50 mg · kg

Objective To study the trend of stroke mortality in Minhang District,Shanghai from 2012 to 2022 and to predict stroke mortality from 2023 to 2027.Methods Annual percentage change(APC)of stroke deaths in Minhang District,Shanghai from 2012 to 2022 was calculated,and then Joinpoint linear regression model was used to analyze the time trend of stroke deaths.A grey GM(1,1)model was constructed based on the stroke mortality rate in Minhang District,Shanghai from 2012 to 2022.The model was used to predict and analyze the stroke mortality rate in Minhang District,Shanghai from 2023 to 2027.The fitting effect of the model was evaluated using relative error and grade deviation.Results From 2012 to 2022,the overall mortality rate of stroke in Minhang District,Shanghai was on the rise for both males and females(total population:APC=2.50%,P<0.001;male:APC=3.41%,P<0.001;female:APC=1.46%,P=0.008).The grey GM(1,1)model was used to predict the increasing trend of stroke mortality rate in Minhang District from 2023 to 2027.The crude mortality rate of stroke in the entire population in 2027 would be 97.55/100000,with 112.31/100000 for males and 83.33/100000 for females.The fitting effect of the model was tested and evaluated to meet high requirements.Conclusion In the past decade,the mortality rate of stroke in Minhang District,Shanghai has shown a significant upward trend.The 5-year prediction results showed that the mortality rate will still on the rise year by year.

Periodontitis is a common chronic inflammatory disease that causes the periodontal bone destruction and may ultimately result in tooth loss.With the progression of periodontitis,the osteoimmunology microenvironment in periodontitis is damaged and leads to the formation of pathological alveolar bone resorption.CD301b+macrophages are specific to the osteoimmunology microenvironment,and are emerging as vital booster for conducting bone regeneration.However,the key upstream targets of CD301b+macrophages and their potential mechanism in periodontitis remain elusive.In this study,we concentrated on the role of Tim4,a latent upstream regulator of CD301b+macrophages.We first demonstrated that the transcription level of Timd4(gene name of Tim4)in CD301b+macrophages was significantly upregulated compared to CD301b-macrophages via high-throughput RNA sequencing.Moreover,several Tim4-related functions such as apoptotic cell clearance,phagocytosis and engulfment were positively regulated by CD301b+macrophages.The single-cell RNA sequencing analysis subsequently discovered that Cd301b and Timd4 were specifically co-expressed in macrophages.The following flow cytometric analysis indicated that Tim4 positive expression rates in total macrophages shared highly synchronized dynamic changes with the proportions of CD301b+macrophages as periodontitis progressed.Furthermore,the deficiency of Tim4 in mice decreased CD301b+macrophages and eventually magnified alveolar bone resorption in periodontitis.Additionally,Tim4 controlled the p38 MAPK signaling pathway to ultimately mediate CD301b+macrophages phenotype.In a word,Tim4 might regulate CD301b+macrophages through p38 MAPK signaling pathway in periodontitis,which provided new insights into periodontitis immunoregulation as well as help to develop innovative therapeutic targets and treatment strategies for periodontitis.

Purpose::To identify the risk factors for training-related lower extremity muscle injuries in young males by a non-invasive method of body composition analysis.Methods::A total of 282 healthy young male volunteers aged 18 -20 years participated in this cohort study. Injury location, degree, and injury rate were adjusted by a questionnaire based on the overuse injury assessment methods used in epidemiological studies of sports injuries. The occurrence of training injuries is monitored and diagnosed by physicians and treated accordingly. The body composition was measured using the BodyStat QuadScan 4000 multifrequency Bio-impedance system at 5, 50, 100 and 200 kHz to obtain 4 impedance values. The Shapiro-Wilk test was used to check whether the data conformed to a normal distribution. Data of normal distribution were shown as mean ± SD and analyzed by t-test, while those of non-normal distribution were shown as median (Q 1, Q 3) and analyzed by Wilcoxon rank sum test. The receiver operator characteristic curve and logistic regression analysis were performed to investigate risk factors for developing training-related lower extremity injuries and accuracy. Results::Among the 282 subjects, 78 (27.7%) developed training injuries. Lower extremity training injuries revealed the highest incidence, accounting for 23.4% (66 cases). These patients showed higher percentages of lean body mass ( p = 0.001), total body water (TBW, p=0.006), extracellular water ( p=0.020) and intracellular water ( p=0.010) as well as a larger ratio of basal metabolic rate/total weight ( p=0.006), compared with those without lower extremity muscle injuries. On the contrary, the percentage of body fat ( p=0.001) and body fat mass index ( p=0.002) were lower. Logistic regression analysis showed that TBW percentage > 65.35% ( p=0.050, odds ratio =3.114) and 3rd space water > 0.95% ( p=0.045, odds ratio =2.342) were independent risk factors for lower extremity muscle injuries. Conclusion::TBW percentage and 3rd space water measured with bio-impedance method are potential risk factors for predicting the incidence of lower extremity muscle injuries in young males following training.

The aim of our research was to obtain Listeria bacteriophages from food and related environments,and conduc-ted the analysis of the electron microscopic morphology,host range specificity,and biological characteristics of the purified phages.The double-layer agar method and the spot test were employed for the isolation and identification of a virulent Listeria phage named LMLPA5,with the isolated strain Listeria in-nocua Lin08 as the host.Phage morphology was observed by transmission electron microscope.The biological characteris-tics of the phage were assessed by determining their host range,optimal multiplicity of infection(MOI),one-step growth curve,and physicochemical stability.Additionally,the preservation efficacy of the phage at 4 ℃,-20 ℃,and-80 ℃ was explored.The phage LMLPA5 belongs to the family Myoviridae based on morphology,exhibiting clear and transparent plaques without halo surrounded.Strains of sever-al Listeria species and different serotypes strains of Listeria monocytogenes were susceptible to lysis by LMLPA5,indica-ting its broad-spectrum activity against Listeria monocytogenes.Optimal MOIs and single-step growth curve analyses revealed optimal MOIs of 0.1 and latent period of 10 minutes for LMLPA5,with average burst size at 95.2 PFU/cell.LMLPA5 was sensitive to high temperatures,and completely inactivated after exposure to 70 ℃ for 1 h,while the phage remained stable for over 32 hours ranging from 4 ℃ to 40 ℃.Within the pH range of 4 to 10,phage titer remained stable and completely inactiva-ted until 60 minutes of ultraviolet exposure.LMLPA5 displayed insensitivity to chloroform,confirming its non-enveloped phage morphology.The phages remained stable for over 8 months when store at 4 ℃ and-80 ℃.The biological characteristics and lysis capacity of phage LMLPA5 were elucidated in this study,which provide the basis for further application.

Objective To explore the effect of decoction of Euphorbia fischeriana Steud.and jujuba(DEFSJ)against estrogen receptor(ER)negative(-)and ER positive(+)breast cancer via the PI3k/Akt pathway,and to provide a reference for the targeted treatment of breast cancer.Methods DEFSJ extract was prepared and analyzed using UHPLC-Triple Quad.DEFSJ containing serum(CS)was prepared via a serum pharmacology method.Different concentrations of DEFSJ-CS were applied to(ER-)MDA-MB-453 and(ER+)MCF-7 breast cancer cells in vitro for 48 h.The distribution of cells in different stages of the cellular cycle was evaluated using a Flow cytometer.DNA ladder assay was used to assess the degree of apoptosis,and the expression of PI3k/Akt pathway-related proteins was evaluated by Western blot assay.The expression of FoxO3a,FoxOla,and Bim mRNA was detected by Real-time quantitative PCR.Nuclear transposition of FoxO3a protein was analysed using a confocal laser microscopy.Results Five batches of DEFSJ extract were analyzed using UPLC,and the result showed that the preparation technology was feasible and the quality was controllable,ensuring the accuracy of the pharmacological experiment result.DEFSJ-CS blocked cells in the G2/M phase(P＜0.05,P＜0.01).Cells treated with DEFSJ-CS displayed the typical apoptotic ladder in the DNA ladder experiment.Compared with the negative control cells,the DEFSJ-CS group cells had decreased protein expression of p-PI3k,p-Akt,p-FoxO3a,and p-FoxO1a(P＜0.05,P＜0.01);increased protein expression of Bim(P＜0.05);decreased mRNA expression of FoxO3a and FoxO1a(P＜0.05,P＜0.01);increased mRNA expression of Bim(P＜0.05,P＜0.01);and enhanced nuclear translocation of FoxO3a protein.The data showed that DEFSJ-CS had a stronger effect on(ER-)MDA-MB-453 cells than(ER+)MCF-7 cells.Conclusions The regulatory effect of DEFSJ extract on anti-breast cancer involves the PI3k/Akt pathway,and the effect varies with phenotypic differences.