The purified 3C8 was obtained by two step column purification,including Protein G affinity purification and DEAE anion exchange purification.The purity of purified 3C8 was about 93％ when analyzed by reverse column.SDS-PAGE showed that the purity of 3C8 was increased greatly by two step purification.By flowcytometry we found that 3C8 specifically binded with B7-H4/293T cells and did not bind with Mock/293T cells,moreover 3C8 did not bind with other B7 family members transgene cells.In confocal experiment 3C8 could specifically stained B7-H4/293T cells.In Western blot only B7-H4/293T cells showed positive band while Mock/293T cells showed negative result.The result of immunohistochemistry showed that B7-H4 was highly expressed in prostate cancer and renal cell carcinoma,while para-cancer tissues did not express B7-H4.The T cell proliferation experiment showed that B7-H4-Ig could bind to activate T cells and inhibit T cell proliferation,while 3C8 could block the binding of B7-H4-Ig and reverse the T cell proliferation inhibition effect of B7-H4-Ig by CFSE and CCK8 assay.The cytokine IFN-γ and IL-2 secreted by activating T cells was decreased by B7-H4-Ig and 3C8 could reverse the effect of B7-H4-Ig.

Objective To investigate doctor's screening practice for hepatitis B virus (HBV) infection before immunosuppressive therapy for rheumatoid arthritis (RA) patients and clinical management of RA patients with positive surface antigen of HBV (HBsAg).Methods One hundred fifty doctors who treated RA patients in daily clinic were survied with a modified American College of Rheumatology (ACR) questionnaire which was composed of demographic data and 10 multiple-choice questions.Step-forward logistic regression analysis was performed to find out the influencing factors,then receiver operator characteristic curve analysis and area under the curve were performed to confirm the influencing factors.Results One hundred and thirtytwo effective questionnaires were collected.Before immunosuppressive therapy,HBV screening rate in outpatients with RA was significandy lower than that in hospitalized patients (68.7％ vs 94.6％,x2=31.5,P＜0.01).Only 23.7％(31/131) of doctors considered antiviral treatment for all RA patients with positive HBsAg.One hundred and thirteen doctors had clinical experience of antiviral treatment,but only 30.1％(34/113) and 23.9％ (27/113) of these doctors chose entecavir or adefovir as the antiviral drug respectively,59.3％ (67/113) prescribed antiviral drug before or together with immunosuppressive therapy compared with 40.7％(46/113) after HBV reactivation.Only 20.4％(23/113) of doctors would sustain antiviral treatment until the termination of steroid or disease modifying antirheumatic drugs (DMARDs).During immunosuppressive therapy for HBsAg(+) RA patients,11.4％(15/132) and 30.3％(40/132) of doctors reported no regular monitoring of aminotransferase or HBV DNA respectively.Conclusion Our survey shows that HBV screening rate in outpatients with RA is low and low awareness of antiviral treatment for all RA patients with positive HBsAg,and lack of awareness of indication,choosing of antiviral drugs,initiation,monitoring and duration of antiviral treatment during immunosuppressive therapy.Further medical education on the associated information and importance to collaborate with hepatologists should be emphasized.

Objective We evaluate if supplementary grading system can refine patient selection and accurately predict neurological outcome in BAVM. Methods We retrospectively study 221 BAVM patients who were treated micro?surgically by our hospital. The score of pre and post operation mRS and relative clinical, radiology data were collected. Two different logistic models (Spetzler-Martin, Supplementary Spetzler-Martin grading model) were constructed to com?pare the area under ROC. Results Some factors are significant different between worse outcome patients and good out?come patients:Non-hemorrhagic presentations prior surgery, AVM bigger than 3cm, diffuse shape of AVM and the elder patients. Predictive accuracy was higher for the supplementary model (ROC area, 0.91), than the Spetzler-Martin model (ROC area, 0.774). So the predictive accuracy of supplementary model was significantly better than that of the Spet?zler-Martin model (P=0.0362). Conclusions Supplementary Spetzler-Martin model can improve preoperative risk pre?diction and subgroup the patients more efficiently. When the score less than 5(including 5) in supplementary Spet?zler-Martin patients seem to have lower risk relative to surgery.

BACKGROUND:As the high proliferation and low apoptosis of the bone marrow in polycythemia vera patients, hematopoietic stem cels transplanted into NOD/SCID mice can differentiate into erythroid cels, but whether hematopoietic stem cels transplantation could improve the hematopoietic function of aplastic anemia mice is not yet reported. OBJECTIVE:To investigate whether transplantation of bone marrow mononuclear cels with JAK2V617F mutation from polycythemia vera patients can influence hematopoietic reconstruction in aplastic anemia mice. METHODS:Severe aplastic anemia mouse models were established by using recombinant human interferon-γplus busulfan, and then, these mouse models were randomly divided into experimental group (n=10) and control group (n=10). Bone marrow mononuclear cels isolated from polycythemia vera patients with positive JAK2V617F mutation were transplanted into the mice in the experimental group via tail vein at 5 days after drug withdrawal.The same volume of normal saline was administered to the control group. Routine peripheral blood test, morphology of bone marrow cels, bone marrow biopsy, and percentage of CD45+ cels in the peripheral blood and marrow were determined at 14 days after transplantation. RESULTS AND CONCLUSION: At 14 days after transplantation, pancytopenia occurred in the experimental group, bone marrow smears showed scattered lymphocytes and hematopoietic progenitors, and bone marrow biopsy presented that hematopoietic tissues were reduced and a smal amount of granulocyte cels and erythroblasts could be seen, but megakaryocytes were rare. In contrast to the control group, there was no improvement in the hematopoietic function of mice in the experimental group. CD45+ cels were detectable in the peripheral blood and bone marrow in the experimental group, but not in the control group; and a higher percentage of CD45+ cels was measured in the bone marrow than in the peripheral blood of experimental group mice. Experimental findings indicate that bone marrow mononuclear cels from polycythemia vera patients with positive JAK2V617F mutation can be engrafted into aplastic anemia mice, but cannot improve the hematopoietic function of mice.

Objective:To explore the different coagulation state in patients with adenomyosis and its clinical significance.Methods:Clinical data of the patients admitted to the First Affiliated Hospital of Nanjing Medical University from January 2017 to December 2021 were retrospectively analyzed. (1) Differential coagulation state between 25 healthy women and 25 patients with adenomyosis were compared during menstrual and non-menstrual periods. (2) The coagulation indexes of 145 patients with adenomyosis (observation group 1) and 129 patients with cervical intraepithelial neoplasia grade Ⅲ (control group 1) who underwent hysterectomy in non-menstrual period were compared. (3) The coagulation indexes of 154 patients with adenomyosis (observation group 2) and 147 women without myometrial lesions (control group 2) who underwent endometrial curettage during uterine bleeding period were compared. (4) Correlations of coagulation index with cancer antigen 125 (CA 125), cancer antigen 19-9 (CA 19-9) and uterine volume in patients with adenomyosis were analyzed. Results:(1) The coagulation state of each health women during the menstrual and non-menstrual period showed no significant differences (all P>0.05). For the 25 patients with adenomyosis, fibrinogen [FIB; 2.61 g/L(2.50-3.10 g/L)] and D-dimer [0.60 mg/L (0.40-1.00 mg/L)] in the menstrual period were significantly higher than those in the non-menstrual period [2.25 g/L (1.90-2.70 g/L) and 0.27 mg/L (0.20-0.40 mg/L), respectively; both P<0.01], while thrombin time [TT; 16.70 s (16.10-17.40 s)] in the menstrual period was significantly lower than that in the non-menstrual period [17.95 s (17.20-18.40 s); P<0.01]. (2) In the non-bleeding period, D-dimer [0.26 mg/L (0.20-0.40 mg/L)] and platelet count [257.0×10 9/L (212.0×10 9/L-308.5×10 9/L)] of observation group 1 were significantly higher than those of control group 1 (all P<0.01). Besides, FIB ( r=0.237, P=0.004) and D-dimer ( r=0.373, P<0.001) were positively correlated with CA 125, while prothrombin time (PT; r=-0.208, P=0.012) and internationalized normalized ratio of plasma prothrombin time (PT-INR; r=-0.201, P=0.015) were negatively correlated with CA 19-9. (3) In the bleeding period, PT [10.70 s (10.10-11.20 s)] and PT-INR [0.93 (0.90-1.00)] of observation group 2 were significantly lower than those of control group 2 (all P<0.01), while D-dimer [0.41 mg/L (0.20-0.80 mg/L)] was significantly higher than that in the control group 2 ( P<0.001). Furthermore, FIB ( r=0.252, P=0.038) and D-dimer ( r=0.321, P=0.008) were positively correlated with uterine volume, while activated partial thromboplastin time (APTT; r=-0.190, P=0.018) and TT ( r=-0.304, P=0.012) were negatively correlated with uterine volume. (4) During non-menstrual period and uterine bleeding period, APTT and TT in patients of observation group 1 and 2 combined with anemia were significantly lower than those of non-anemia patients (all P<0.05). Conclusion:Patients with adenomyosis have a tendency to hypercoagulability in both the uterine bleeding and non-bleeding periods, which may be related to enlarged uterine volume, increased serum CA 125 and anemia.

Objective:To evaluate the value of preoperative prediction of vessel invasion (VI) of locally advanced gastric cancer by machine learning model based on the venous phase enhanced CT radiomics features.Methods:A retrospective analysis of 296 patients with locally advanced gastric cancer confirmed by pathology in the First Affiliated Hospital of Zhengzhou University from July 2011 to December 2020 was performed. The patients were divided into VI positive group ( n=213) and VI negative group ( n=83) based on pathological results. The data were divided into training set ( n=207) and test set ( n=89) according to the ratio of 7∶3 with stratification sampling. The clinical characteristics of patients were recorded, and the independent risk factors of gastric cancer VI were screened by multivariate logistic regression. Pyradiomics software was used to extract radiomic features from the venous phase enhanced CT images, and the minimum absolute shrinkage and selection algorithm (LASSO) was used to screen the features, obtain the optimal feature subset, and establish the radiomics signature. Four machine learning algorithms, including extreme gradient boosting (XGBoost), logistic, naive Bayes (GNB), and support vector machine (SVM) models, were used to build prediction models for the radiomics signature and the screened clinical independent risk factors. The efficacy of the model in predicting gastric cancer VI was evaluated by the receiver operating characteristic curve. Results:The degree of differentiation (OR=13.651, 95%CI 7.265-25.650, P=0.003), Lauren′s classification (OR=1.349, 95%CI 1.011-1.799, P=0.042) and CA199 (OR=1.796, 95%CI 1.406-2.186, P=0.044) were independent risk factors for predicting the VI of locally advanced gastric cancer. Based on the venous phase enhanced CT images, 864 quantitative features were extracted, and 18 best constructed radiomics signature were selected by LASSO. In the training set, the area under the curve (AUC) of XGBoost, logistic, GNB and SVM models for predicting gastric cancer VI were 0.914 (95%CI 0.875-0.953), 0.897 (95%CI 0.853-0.940), 0.880 (95%CI 0.832-0.928) and 0.814 (95%CI 0.755-0.873), respectively, and in the test set were 0.870 (95%CI 0.769-0.971), 0.877 (95%CI 0.788-0.964), 0.859 (95%CI 0.755-0.961) and 0.773 (95%CI 0.647-0.898). The logistic model had the largest AUC in the test set. Conclusions:The machine learning model based on the venous phase enhanced CT radiomics features has high efficacy in predicting the VI of locally advanced gastric cancer before the operation, and the logistic model demonstrates the best diagnostic efficacy.

Objective: To evaluate the effectiveness of reactive oxygen species (ROS)-responsive nanomedicine in suppressing corneal neovascularization (CNV) in vivo. Methods: ROS-responsive nanomedicine (ROS-TK-5/siVEGF), which consists of vascular endothelial growth factor (VEGF) small interfering RNA (siRNA) and thioketal linkage was synthesized by the Michael addition.The cumulative release of siVEGF from nanomedicine under oxidant conditions was assessed by agarose gel electrophoresis.Thirty-nine VEGFR2-luc-KI transgenic mice were used in this study, of which 30 mice were randomly divided into a normal control group, a PBS control group, an ROS-TK-5/NC group, an ROS-TK-5/siVEGF group, and a ranibizumab group, with 6 mice in each group.The ROS levels in the corneal tissue after alkali burning were tested by dihydroethidium (DHE) staining in the other 9 mice.In each group, alkali-burned mice were subconjunctivally injected with 10 μl of a different formula every two days.The effectiveness of nanomedicine in attenuating CNV was evaluated by slit-lamp microscopy and an in vivo imaging system (IVIS) at 7, 14, and 21 days after alkali burning. The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology (ARVO) and the Guidelines of the Animal Experimental Committee of Liberation Army General Hospital.The study protocol was approved by the Ethics Committee of Liberation Army General Hospital (No.2018-X14-82). Results: After treathrent with an aqueous solution without ROS, only 5%-10% of the siVEGF was released from the nanoparticles within 10 hours.In contrast, about 70% of the siVEGF was released from the nanoparticles after treatment with 10 mmol/L H₂O₂ within 10 hours.The relative fluorescent intensities in the corneal stromal layer at 7 days and 14 days after alkali burning were 5.403±0.306 and 2.930±0.255, respectively, which was significantly greater than those in the normal control group (1.003±0.015) (both at P<0.05). The CNV areas were statistically different among the four groups at various time points (F_group=49.855, P<0.01; F_time=65.556, P<0.01). The CNV area was significantly reduced in the ROS-TK-5/siVEGF and ranibizumab groups compared with the PBS control and ROS-TK-5/NC groups at 7 days and 14 days after modeling, and the CNV area was more effectively reduced in the ROS-TK-5/siVEGF group than the ranibizumab group at 7 days and 14 days after modeling (all at P<0.05). At day 21 after modeling, the CNV area was significantly reduced in the ROS-TK-5/siVEGF and ranibizumab groups compared to the PBS control and ROS-TK-5/NC groups (all at P<0.05). IVIS showed that the corneal fluorescent intensity was statistically different among the four groups at various times (F_group=27.193, P=0.003; F_time=51.062, P<0.01). The corneal fluorescent intensities were significantly reduced in the ROS-TK-5/siVEGF and ranibizumab groups compared to the PBS control and ROS-TK-5/NC groups at 7 days and 14 days after modeling; in addition, the corneal fluorescent intensity was more effectively reduced in the ROS-TK-5/siVEGF group in comparison with the ranibizumab group at 7 days and 14 days after modeling (all at P<0.05). At 21 days after modeling, the corneal fluorescent intensity was significantly reduced in the ROS-TK-5/siVEGF and ranibizumab groups compared to the PBS control and ROS-TK-5/NC groups (all at P<0.05). Conclusions ROS-TK-5/siVEGF nanomedicine effectively attenuates alkali burn-induced CNV formation and appears to have a better effect in comparison with ranibizumab at an early stage.

Objective:To describe the study methods and baseline characteristics of participants enrolled in mCessation program.Methods:This is a longitudinal, real-world study with non-randomized controlled design. The mCessation program consisted of a WeChat official account, an applet and a website using the same name ‘mCessation Online’. After users followed the WeChat account, filled in baseline information online and set a quit date, they would receive 162 short text messages in the next six and a half months as scheduled. This study collected the information of participants enrolled from May 26, 2021 to September 30, 2022, and analyzed baseline data including demographic characteristics, smoking characteristics, degree of tobacco dependence, reasons for smoking cessation and other related factors.Results:During the study period, a total of 16 746 participants registered, and 13 887 participants (82.9%) were enrolled in final analysis after screening the inclusion and exclusion criteria and completion of main indicators. Each year the number of enrolled participants in May or June was 1 381 to 2 707 per month, higher than the number of enrolled participants in other months (233 to 569 per month). Participants from North China accounted for the largest proportion (29.3%). There were 13 316 men (95.9%) in enrolled participants and the mean age was (36±10) years. Most participants were 25-34 (38.8%) or 35-44 (30.8%) years old. In terms of smoking characteristics, there were 12 564 (90.5%) daily smokers. The starting age of smoking was 18 (15, 20) years old. 11 866 participants (85.4%) were tobacco dependent, mostly with degree of mild (76.4%) or moderate (20.2%). In terms of reasons for quitting, 9 315 participants′ (67.1%) reasons were to prevent disease, 6 742 participants (48.5%) were concerned about impact of smoking on family members, and 6 731 participants (48.5%) were under requested by families.Conclusion:mCessation program can effectively recruit smokers with intention to quit in short time, especially those who were male, young and tobacco dependent.