Objective To extract hyperoside from the leaves of Acanthopanax sessiliflorus by complex enzyme method, and optimize the extraction process by orthogonal experiment. Methods Hyperoside was determined by HPLC. Effects of temperature,α-amylase, neutral protease and cellulase on extraction rate were detected by the orthogonal tests, and the optimum extraction condition of hyperoside from the leaves of Acanthopanax sessiliflorus was determined by complex enzyme method. Results The main influence factor was temperature,follows byα-amylase, neutral protease and cellulase according to orthogonal analysis.The best condition was as follows: dose of cellulase, neutral protease and α-amylase was 2%, 0. 5% and 3%, respectively, extract at temperature of 30 ℃for 10 min. Under this condition, the extraction rate of hyperoside in the leaves of Acanthopanax senticosus was 0.52%. Conclusion As compared with the traditional technics, compound enzyme increases the productivity of hyperoside.

Objective To investigate the correlations of serum Apelin-13 and fatty acid binding protein 4(FABP4)levels with metabolic and bone metabolic indicators in postmenopausal women with different bone mass.Methods A total of 145 postmenopausal women were selected as subjects and divided into three groups based on bone mineral density(BMD)test results:normal bone mass group(49 cases),osteopenia(ON)group(51 cases),and osteoporosis(OP)group(45 cases).Se-rum Apelin-13,FABP4 levels,bone metabolic indicators,and biochemical indicators were measured and compared among the three groups.Spearman correlation analysis was used to analyze the correla-tions of Apelin-13,FABP4,and other indicators with BMD.Multivariate Logistic regression analysis was performed to analyze the risk factors for OP,and the receiver operating characteristic(ROC)curve was plotted to analyze the predictive value of serum Apelin-13 for postmenopausal osteoporosis(PMOP).Results The serum Apelin-13 level in the OP group was lower than that in the ON group and the normal bone mass group(P＜0.05).No significant difference in serum FABP4 levels was found among the three groups(P＞0.05).The levels of serum parathyroid hormone(PTH),alkaline phosphatase(ALP),type Ⅰ procollagen amino-terminal propeptide(PⅠNP),type Ⅰ collagen cross-linked C-terminal peptide(CTX Ⅰ),and bone-specific alkaline phosphatase(BALP)in the OP group were higher than those in the ON group and the normal bone mass group(P＜0.05).Lumbar post-menopause BMD was positively correlated with serum Apelin-13 levels(P＜0.05),but had no cor-relation with serum FABP4 levels(P＞0.05).Lumbar BMD was negatively correlated with serum PTH,ALP,P Ⅰ NP,CTX Ⅰ,BALP,and age(P＜0.05),but positively correlated with body weight,body mass index,T-score,fasting insulin,and insulin resistance index(P＜0.05).Multi-variate Logistic regression analysis showed that serum Apelin-13,PTH,ALP,P Ⅰ NP,CTX Ⅰ,and BALP levels were independent factors influencing the occurrence of OP in postmenopausal women(P＜0.05).ROC curve results showed that the optimal cut-off value of serum Apelin-13 for predic-ting PMOP was 18.51 pg/mL,with an area under the curve of 0.716,a sensitivity of 70.0％,and a specificity of 64.4％.Conclusion Apelin-13 is lowly expressed in the serum of PMOP patients,and its expression level is closely related to lumbar BMD,which may serve as an early screening in-dicator and potential therapeutic target for PMOP.

Objective To investigate the correlations of serum Apelin-13 and fatty acid binding protein 4(FABP4)levels with metabolic and bone metabolic indicators in postmenopausal women with different bone mass.Methods A total of 145 postmenopausal women were selected as subjects and divided into three groups based on bone mineral density(BMD)test results:normal bone mass group(49 cases),osteopenia(ON)group(51 cases),and osteoporosis(OP)group(45 cases).Se-rum Apelin-13,FABP4 levels,bone metabolic indicators,and biochemical indicators were measured and compared among the three groups.Spearman correlation analysis was used to analyze the correla-tions of Apelin-13,FABP4,and other indicators with BMD.Multivariate Logistic regression analysis was performed to analyze the risk factors for OP,and the receiver operating characteristic(ROC)curve was plotted to analyze the predictive value of serum Apelin-13 for postmenopausal osteoporosis(PMOP).Results The serum Apelin-13 level in the OP group was lower than that in the ON group and the normal bone mass group(P＜0.05).No significant difference in serum FABP4 levels was found among the three groups(P＞0.05).The levels of serum parathyroid hormone(PTH),alkaline phosphatase(ALP),type Ⅰ procollagen amino-terminal propeptide(PⅠNP),type Ⅰ collagen cross-linked C-terminal peptide(CTX Ⅰ),and bone-specific alkaline phosphatase(BALP)in the OP group were higher than those in the ON group and the normal bone mass group(P＜0.05).Lumbar post-menopause BMD was positively correlated with serum Apelin-13 levels(P＜0.05),but had no cor-relation with serum FABP4 levels(P＞0.05).Lumbar BMD was negatively correlated with serum PTH,ALP,P Ⅰ NP,CTX Ⅰ,BALP,and age(P＜0.05),but positively correlated with body weight,body mass index,T-score,fasting insulin,and insulin resistance index(P＜0.05).Multi-variate Logistic regression analysis showed that serum Apelin-13,PTH,ALP,P Ⅰ NP,CTX Ⅰ,and BALP levels were independent factors influencing the occurrence of OP in postmenopausal women(P＜0.05).ROC curve results showed that the optimal cut-off value of serum Apelin-13 for predic-ting PMOP was 18.51 pg/mL,with an area under the curve of 0.716,a sensitivity of 70.0％,and a specificity of 64.4％.Conclusion Apelin-13 is lowly expressed in the serum of PMOP patients,and its expression level is closely related to lumbar BMD,which may serve as an early screening in-dicator and potential therapeutic target for PMOP.

Objective To investigate the role and mechanism of sine oculis homeobox 1(Six1),a nephrogenic transcription factor,in regulating injury repair after acute kidney injury(AKI).Methods C57BL/6J mice were inflicted with renal ischemia reperfusion(IR)injury to establish AKI model,and then the serum samples and kidney tissues were collected at days 1,2,and 3 after modeling.Serum creatinine(Cr)and blood urea nitrogen(BUN)levels were measured and renal morphology was observed with HE staining.The expression and distribution of nephrogenic molecules(Six1 and Pax2,etc.)and cell proliferation/migration genes(Cyclin A1,MMP9,etc.)were detected with RT-PCR,Western blotting,and immunofluorescence assay and immunohistochemical assay.The expression of apoptosis pathway(Bc12,Caspase3,etc.)and cell proliferation related molecules were evaluated in Six1 overexpression/knockout human epithelial cells(HK2)with CoCl2-induced injury.Additionally,after the adeno-associated viruses carrying Six1 overexpression vector were used to overexpress the molecule in the mice,the expression of Six1 and other related molecules were detected after IR injury modeling.Results After renal IR injury,Six1 was significantly activated in epithelial cells,the expression of nephrogenic and cell proliferation/migration molecules(Pax2,Cyclin A1,MMP9,etc.)was obviously up-regulated(P＜0.05),which was positively correlated with Six1 expression,while the proliferation/migration molecules(Ki67,MMP9)were also localized within the tubular epithelial cells.In cellular models of Six1 overexpression,the expression levels of nephrogenic and cell proliferation/migration molecules(Pax2,Cyclin A1,MMP9,etc.)were also notably up-regulated(P＜0.05).In the mice with renal overexpression of Six1,the nephrogenic molecules as well as anti-apoptotic ones(Six2,Pax2,Bc12,etc.)were up-regulated,while the expression of kidney injury-related molecule(Kim1)in kidneys was reduced in the renal tissues.While,in 2 d after IR injury,the anti-apoptotic gene(Bc12,Stat3)was significantly up-regulated(P＜0.05),and the apoptotic and injury molecules(Kim1,Caspase3)showed remarkable down-regulation(P＜0.05)in the mice with renal Six1 overexpression.Furthermore,CoCl2-inducion significantly decreased the cell proliferation rate in the Six1 knockoutgroup(TCMK1-Six1-/-)(P＜0.05)but increased the rate in the Six1 overexpression group(TCMK1-Six1)when compared to the control cells(P＜0.05).And,the expression of nephrogenic,anti-apoptotic pathways and cell proliferation/migration molecules(Pax2,Bc12,Cyclin A1,MMP9,etc.)were reduced in TCMK1-Six1-/-group,and apoptosis and kidney injury molecules(Caspase3,Kim1)were significantly down-regulated in TCMK1-Six1 group(P＜0.05).Conclusion Activation of Six1 after AKI can promote the proliferation/migration of renal tubular epithelial cells by up-regulating nephrogenic molecules and inducing anti-apoptotic pathway molecules,and then,participate in IR-induced renal injury repair.

Objective:To assess the influence of an artificial intelligence (AI) -assisted diagnosis system on the performance of endoscopists in diagnosing gastric cancer by magnifying narrow banding imaging (M-NBI).Methods:M-NBI images of early gastric cancer (EGC) and non-gastric cancer from Renmin Hospital of Wuhan University from March 2017 to January 2020 and public datasets were collected, among which 4 667 images (1 950 images of EGC and 2 717 of non-gastric cancer)were included in the training set and 1 539 images (483 images of EGC and 1 056 of non-gastric cancer) composed a test set. The model was trained using deep learning technique. One hundred M-NBI videos from Beijing Cancer Hospital and Renmin Hospital of Wuhan University between 9 June 2020 and 17 November 2020 were prospectively collected as a video test set, 38 of gastric cancer and 62 of non-gastric cancer. Four endoscopists from four other hospitals participated in the study, diagnosing the video test twice, with and without AI. The influence of the system on endoscopists′ performance was assessed.Results:Without AI assistance, accuracy, sensitivity, and specificity of endoscopists′ diagnosis of gastric cancer were 81.00%±4.30%, 71.05%±9.67%, and 87.10%±10.88%, respectively. With AI assistance, accuracy, sensitivity and specificity of diagnosis were 86.50%±2.06%, 84.87%±11.07%, and 87.50%±4.47%, respectively. Diagnostic accuracy ( P=0.302) and sensitivity ( P=0.180) of endoscopists with AI assistance were improved compared with those without. Accuracy, sensitivity and specificity of AI in identifying gastric cancer in the video test set were 88.00% (88/100), 97.37% (37/38), and 82.26% (51/62), respectively. Sensitivity of AI was higher than that of the average of endoscopists ( P=0.002). Conclusion:AI-assisted diagnosis system is an effective tool to assist diagnosis of gastric cancer in M-NBI, which can improve the diagnostic ability of endoscopists. It can also remind endoscopists of high-risk areas in real time to reduce the probability of missed diagnosis.

Objective:To prepare a novel tri-specific T cell engager (19TriTE) targeting CD19 antigen, and to investigate its immunotherapeutic effect on CD19-positive hematological malignancies.Methods:19TriTE was constructed by molecular cloning technology and successfully expressed through the eukaryotic expressing system. The effects of 19TriTE on the proliferation and activation of T cells, as well as the specific cytotoxicity against CD19 positive tumor cell lines were verified.Results:①19TriTE expressing plasmid was constructed and successfully expressed through the eukaryotic expressing system. ②19TriTE can specifically bind to T cells and Nalm6 cells, with equilibrium dissociation constants of 19.21 nmol/L and 11.67 nmol/L, respectively. ③The expression rates of CD69 positive T cells and CD25 positive T cells were 35.4% and 49.8% respectively, when 2 nmol/L 19TriTE were added in the co-culture system, which were significantly higher than those in the control group. ④19TriTE can significantly promote the proliferation of T cells. The absolute count of T cells expanded from the initial one million to 74 million with an 74 fold increase at the concentration of 1 nmol/L on day 12. ⑤19TriTE can significantly mediate T cells killing of CD19 positive target cells in a dose-dependent manner. At the concentration of 10 nmol/L, the target cells lysis reached 50%. ⑥Degranulation experiment verified that 19TriTE can activate T cells in the presence of CD19 positive target cells, and the activation of T cells positively correlated with the dose of 19TriTE. ⑦When 19TriTE fusion protein co-cultured with T cells and target cells overexpression RFP and luciferase genes respectively, 19TriTE can notably mediate T cells killing of CD19 positive target cells through fluorescent microscope or bioluminescence imaging technology.Conclusion:In this study, we successfully constructed and expressed 19TriTE fusion protein and verified that it can effectively activate T cells and promote their proliferation in vitro. At the same time, it can bind to CD19 positive target cells and T cells, as well as enhance T cells anti-leukemia effect in vitro, providing the foundation for further clinical research.