Objective We aimed to investigate the hemostatic effect and mechanism of modified Sijunzi Decoction(MSJZT),consisting of tangshen,India bread,largehead atractylodes rhizome,liquorice root,milkvetch root,ass hide glue,and India madder root,on the zebrafish intestinal bleeding model through 5-hydroxytryptamine(5-HT)and its receptors(5-HT2aR,5-HT2bR).Methods Zebrafish at 4 days post fertilization were used as the research object.An intestinal bleeding model was established by induction with 6 μmol/L simvastatin for 24 hours.The zebrafish were divided into normal group,model group,MSJZT low-,mid-and high-concentration groups(55.6,167,500 mg/L),and after modeling,the corresponding concentration of MSJZT was administered.The samples were collected after 24 hours.Platelet count(PLT)and bleeding status were observed.The content of 5-HT and Caspase 3/7 activity were detected by colorimetry.The gene expressions of 5-HT2aR,5-HT2bR and serotonin transporter(SERT)were detected by real-time PCR.The protein expressions of protein kinase B(AKT),phosphorylated AKT(p-AKT),extracellular signal-regulated protein kinase(ERK),and phosphorylated ERK(p-ERK)were detected by Western blotting.Results The intestinal bleeding rate in the model group was 70.0%,and that in the MSJZT low-,mid-and high-concentration groups was 36.7%,40.0%,and 80.0%,respectively;the intestinal hemostatic effect was 54%,52%,and 7%,respectively.Compared with the model group,the PLT in the MSJZT low-and mid-concentration groups decreased;the content of 5-HT in all MSJZT groups increased,and the gene expressions of 5-HT2aR and 5-HT2bR were up-regulated.The gene expression of SERT was up-regulated in the low-and mid-concentration groups;the Caspase 3/7 activity in the mid-and high-concentration groups was reduced;the expressions of p-AKT and p-ERK proteins were up-regulated in the low-and mid-concentration groups(all P<0.05).Conclusion MSJZT can reduce the incidence and severity of intestinal bleeding in zebrafish intestinal bleeding models.The mechanism may be achieved by activating AKT and ERK signaling pathways through 5-HT and its receptors.

Objective To investigate the clinical efficacy and safety of pomegranate blood replenishing syrup in the treatment of iron deficiency anemia.Methods Using a multicenter,randomized,open,positive drug-controlled method,120 patients with iron deficien-cy anemia of the qi and blood deficiency type admitted from June 1,2020 to March 1,2023 were divided into the treatment group(n=90)and the control group(n=30),with pomegranate blood replenishing syrup was given to the patients in treatment group and shengshu-bao combination was given to the patients in control group.The clinical efficacy,Chinese medicine evidence and adverse reactions were compared between the two groups.Results The effective rate of the treatment group was 52.22％,which was better than 30％of the con-trol group,and the difference was statistically significant(P＜0.05).The hemoglobin level of the treatment group was higher than that of the control group after 8 weeks of treatment,and the difference was statistically significant(P＜0.05);in terms of TCM syndrome effica-cy,the total effective rate of the treatment group was 82.22％,which was better than 40.00％of the control group,and the difference was statistically significant(P＜0.05).There were no adverse events in either group.Conclusion For patients with iron deficiency anemia of the qi and blood deficiency type,pomegranate blood replenishing syrup has obvious clinical efficacy and good clinical safety.

Objective:To explore the hemostatic mechanism of Jianpi Yiqi Shexue decoction(JYSD)by regulating vascular factors in an immune thrombocytopenia(ITP)mouse model.Methods:An ITP mouse model was established by the passive-immune modeling method,and inter-ventional drugs used were prednisone tablets and JYSD.The platelet count;vascular activity-related factors vWF,VCAM-1,and TM;and VEGF and bFGF were used as observational indicators.Results:On the 8th day of administration,compared with the model group,platelet counts in the prednisone and JYSD groups increased(both P＜.001).Compared with the control group,the levels of vWF,VCAM-1,and TM in the other groups were lower(all P＜.05).The VCAM-1 level in the JYSD group was higher than that in the prednisone group(P=.012),but without significant difference compared with the model group(P=.051).The TM level in the JYSD group was the lowest(vs.the model group,P=.047;vs.the prednisone group,P=.006).Compared with the control group,the IOD values of VEGF and bFGF in the other three groups were lower(all P＜.01).The IOD values of VEGF in the prednisone and JYSD groups were both higher than those in the model group(P=.002 and P＜.001,respectively).The IOD values of bFGF among the model,prednisone,and JYSD groups were not statistically significant(P＞.05).Conclusion:A vascular factor disorder is involved in the pathogenesis of ITP.JYSD can increase the platelet count,upregulate VEGF expression,and reduce the TM level.JYSD has the same effect as prednisone tablets in regulating platelet,vWF,VEGF,and bFGF,with a stronger effect in normalizing VCAM-1 and TM levels.The hemostatic mechanism of JYSD is closely related to the effective balance of vascular factors.

Objective:To explore the application effect of online-offline mixed teaching mode in the teaching of histology and embryology.Methods:Two classes of students majoring in psychiatry from Batch 2020 were randomized into experimental group ( n = 100), and two classes of students majoring in general practice in 2020 grade were randomized into the control group ( n = 99). The online-offline mixed teaching mode was used in the experimental group, while a traditional teaching mode was used in the control group. The online test scores, final scores and questionnaire satisfaction of the two groups of students were evaluated on the teaching effect. All data were statistically processed by SPSS 19.0, and the comparison between groups was performed by t test. Results:The online test scores of the experimental group [(8.55±1.18) vs. (6.33±0.91)] and final scores [(85.56±3.32) vs. (72.24±2.71)] were significantly higher than those of the control group ( P < 0.05). Compared with the control group, the experimental group's satisfaction scores of the online-offline mixed teaching mode were significantly higher in students' learning interest, learning autonomy, time management ability, teamwork, interactive communication, knowledge understanding and memory ability ( P < 0.05). Conclusion:The online-offline mixed teaching mode can improve the histology and embryology teaching effect and promote the students' interest in learning and autonomous learning ability.

OBJECTIVE：To evaluate the effects of nicorand il on the proliferation ，migration ability and Hippo/YAP signaling pathway of pulmonary artery smooth muscle cells （PASMCs）. METHODS ：Human primary PASMCs were divided into normal control group ，model group ，nicorandil low ，medium and high concentration groups （50，100，200 μmol/L），with 3 holes in each group. In addition to the normal control group ，the rest of the cells were inoculated on the gel coated medium to simulate the pulmonary hypertension environment ，so as to establish AS cell model. Then ，each drug group was added with corresponding drugs，and the normal control group and model group were added with the same volume of normal saline ，and cultured for 48 h. CCK-8 assay and Transwell assay were used for the examination of cell proliferation （by light density ）and migration ability ， respectively. mRNA expression of YAP target factors （CTGF and AREG ）were examined by qRT-PCR. Western blotting assay was used to detect the protein expression of CTGF and AREG. RESULTS ：Compared with normal control group ，light density of cells was increased significantly in model group ；the number of migration cells per field of view increased significantly ；mRNA and protein expression of CTGF and AREG were significantly increased （P＜0.01）. Compared with model group ，light density ，the number of migration cells per field of view ，mRNA and protein expression of CTGF and AREG in nicorandil low ，medium and high concentration groups were decreased significantly ， in concentration-dependent manner （P＜0.05 or P＜0.01）. CONCLUSIONS：Nicorandil can inhibit the proliferation and migration of PASMCs in AS model ，the mechanism of which cstc2019jscx-msxmX0174） may be associated with the Hippo/YAP signaling pathway.

Objective:The aim of this study was to study the characteristics of neoplasm invasion type in ovary of two orthotopic models established with human epithelial ovarian cancer solid tumor tissue slices and human ovarian carcinoma cell line OVCAR-3 in nude mice and human epithelial ovarian cancer.Methods:Tumor tissues and cell line OVCAR-3 of human epithelial ovarian cancer were grown in subcutaneous tissue and the subcutaneous tumor source was fetched and inoculated in ovarian capsule of nude mice to establish the orthotopic implantation model. The neoplasm invasion type in the two kinds of models were observed. The neoplasm invasion types were also analyzed by pathological examination in 54 cases of International Federation of Gynecology and Obstetrics (FIGO) stageⅠ-Ⅱepithelial ovarian cancer.Results:Three neoplasm invasion types were found as follows: type of pseudocapsule, type of pseudocapsule invasion, type of pseudocapsule penetration. Pseudocapsule rate in the solid tumor slices group (18.2%) were lower than those in the cell line group (42.3%) ( P<0.05), while the pseudocapsule penetration rate in the solid tumor slices (50.0%) were higher than those in the cell line group (23.1%) ( P<0.05). No difference was found of pseudocapsule invasion rate between two groups ( P>0.05). Neoplasm invasion type in ovary changed with tumor planting time. High proportion of pseudocapsule type was found at the beginning of tumor planting, and the pseudocapsule penetration rate raised with tumor planting time increased. High proportion of pseudocapsule type was also found in patients with FIGO stageⅠepithelial ovarian cancer, and pseudocapsule penetration rate increased in those with FIGO stageⅡ. No difference in neoplasm invasion type was found between two kinds of pathological types ( P>0.05). Conclusions:There are differences between the two kinds of orthotopic models established with human epithelial ovarian cancer solid tumor tissue slices and human ovarian carcinoma cell line OVCAR-3. Compared to the solid tumor slices model, the cell line model is more stable for the follow-up study. The proportion of three neoplasm invasion types in ovary were more balanced in 8 weeks after tumor planting, and 8 weeks after tumor planting is the best start time for the follow-up experiment.

Objective To evaluate the application of Web problem-based learning (WPBL) mode of respiratory system integration curriculum.Methods Experimentclasses 1 and 2 of the 2014 grades of the Qiqihar Medical University were divided into control group(48 students) and experiment group (48 students).Traditional PBL teaching was conducted in control group:the contents of the case were printed and discussed with the material of each class.WPBL teaching was conducted in experiment group:autonomous learning before class,showing cases by Video,asking questions and discussion in group and reporting results in class.The teaching effect was evaluated by PBL evaluation form,questionnaire and final grade.SSPS 19.0 statistical software was used for data analysis and measurement.T test was conducted among groups.Counting data were tested by Chi-square and the results were expressed as percentage.Results The students in the experiment group were better than those of control groupin the aspects of contents understanding,course interest,clinical thinking ability,classroom atmosphere and the course attitude.The scores of PBL (16.65 ±2.82),experiment exam (8.21 ±0.44) and final grade(76.77 ± 12.31) in the experiment group were higher than those in the control group.There are statistical differences in two groups (P＜0.05).Conclusions WPBL teaching can improve students'learning interest,clinical thinking ability and improve the teaching quality of respiratory integration curriculum.

Objective To discuss the application of blending learning based on MOOC in Histology and Embryology, so as to improve students' learning effectiveness. Methods Nursing undergraduates in class one of 2017 grade were randomly selected as experimental group (n=47) , and nursing undergraduates in class two as control group (n=48), with blending learning and traditional teaching model adopted, respec-tively. SPSS 19.0 was applied to processing the data and T test were used to compare the results of the final exam of this course and the evaluation scale of students' independent learning ability in two groups. Results The experimental scores [(24.22±5.08) vs. (18.49±4.65)], paper test scores [(52.37±6.24) vs. (47.15± 5.99)], and total score [(76.61±7.22) vs. (62.83±7.36)], which shows the scores of experimental group were statistically higher than those of the control group. According to the evaluation scale of students' inde-pendent learning ability, the self-learning ability of the experimental group was better than that of the control group, with significant differences (P<0.05). Conclusion Blending learning can be applied to the teaching of Histology and Embryology, which can effectively improve learning quality, with satisfactory teaching effects to both teachers and students.

OBJECTIVE: To explore the genetic basis for a neonate featuring hyperammonemia. METHODS: The patient was examined and tested by tandem mass spectrometry and next generation sequencing (NGS). Suspected mutations were confirmed by Sanger sequencing of the proband and her parents. Potential impact of the mutation was predicted with SIFT, PolyPhen-2 and MutationTaste software. RESULTS: Plasma ammonia and alanine were significantly increased in the proband, while serum citrulline was decreased. The neonate was found to harbor compound heterozygous mutations of the CPS1 gene [c.1631C>T(p.T544M) and c.1981G>T(p.G661C)], which were respectively inherited from her father and mother. CONCLUSION The carbamoyl phosphate synthetase I deficiency of the proband can probably be attributed to the mutations of the CPS1 gene. Above finding has expanded the spectrum of CPS1 mutations in association with carbamoyl phosphate synthetase I deficiency.
Carbamoyl-Phosphate Synthase (Ammonia) ; genetics ; Carbamoyl-Phosphate Synthase I Deficiency Disease ; genetics ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Hyperammonemia ; diagnosis ; genetics ; Infant, Newborn ; Mutation

BACKGROUND:Chemokines can promote (MSCs) the secretion of vasoactive factors from mesenchymal stem cel s (MSCs) through paracrine mechanism, which have important role in accelerating angiogenesis. OBJECTIVE:Under the high glucose environment, to the effect of the supernatant of MSCs stimulated by chemokine (C-X-C motif) ligand 8 (CXCL-8) on human umbilical vein endothelial cel s (HUVECs), and to analyze the mechanism of Sonic Hedgehog signaling pathway in the stimulation of CXCL-8 on MSCs. METHODS:Under the high glucose environment, the MSCs supplemented with 100μg/L CXCL-8 were set as CXCL-8 group;the MSCs that were preprocessed with 5μmol/L octyl maleimide for 45 minutes and then stimulated with 100μg/L CXCL-8 were as Shh inhibitor group;the MSCs that were routinely cultured in a high-glucose medium were as control group. The cel supernatant of each group was extracted as conditioned medium (CM) to culture HUVECs, respectively, and these cel s were referred to as CXCL-8 CM group, Shh inhibitor CM group, and control CM group, respectively. Cel counting kit-8, cel scratch and Transwel chamber tests were used to observe the effect of each CM on HUVEC proliferation, apoptosis and chemotaxis. By establishment of a diabetic skin ulcer model in C57BL/6J mice, the CM of each group was used to treat the mouse model to confirm the effects of CXCL-8 stimulated MSCs CM on HUVEC homing and ulcer healing. RESULTS AND CONCLUSION:(1) The experimental results in vitro:compared with the control CM group, CXCL-8 CM group significantly promoted the proliferation of HUVECs, and decreased the apoptosis of HUVECs, the closure rate and migration rate of HUVECs were significantly increased (P<0.01 or P<0.01), and the levels of vascular endothelial growth factor and epidermal growth factor were significantly increased (P<0.01 or P<0.01). Compared with CXCL-8 CM group, however, the above results in the Shh inhibitor CM group showed reverse changes (P<0.01). (2) The experimental results in vivo:compared with the MSCs CM group and Shh inhibitor CM group, the healing effect of diabetic skin ulcer and the number of HUVECs labeled by green fluorescent protein in the CXCL-8 CM group were significantly increased (P<0.01). To conclude, these findings indicate that CXCL-8 stimulated MSCs secrete paracrine factors, vascular endothelial growth factor and epidermal growth factor, through the Sonic Hedgehog signaling pathway under the high glucose environment, which enhance the homing ability of HUVECs.