Objective: To explore the protective roll of ifbroblast growth factor 21 (FGF21) in endoplasmic reticulum stress (ERS) induced rat's H9c2 cardiomyocyte apoptosis with its mechanism. Methods: pcDNA4 was used as gene vector, pcDNA4-FGF21 plasmid was constructed and transfected into rat's H9c2 myocardiocytes for 48 h. ERS model was established by 10 μM tunicamycin (TM) induction for 24 h. The experiment was conducted in 4 groups:①Control group,②TM group, the cells were treated by TM,③pcDNA4-FGF21+TM group,④pcDNA4+TM group. The expressions of FGF21, protein kinase R-like ER kinase (PERK) and c-Jun N-terminal kinases (JNK) mediated apoptosis pathway related protein were measured by Western blot analysis; cell survival rate was examined by CCK-8 method and apoptosis rate was detected by TUNEL technique. Results: pcDNA4-FGF21 vector was successfully constructed and overexpressed in H9c2 myocardiocytes. Compared with Control group, TM group and pcDNA4+TM group had up-regulated endogenous FGF21 expression, increased PERK and JNK mediated apoptosis pathway related protein expression; reduced cell survival rate and elevated apoptosis rate. Compared with TM group and pcDNA4+TM group, pcDNA4-FGF21+TM group had down-regulated PERK and JNK mediated apoptosis pathway related protein expression; increased cell survival rate and decreased apoptosis rate. Conclusion: FGF21 overexpression can reduce ERS induced apoptosis rat's H9c2 myocardiocytes which might be partly related for inhibiting PERK and JNK mediated signal transduction of apoptosis pathway.

AIM:To explore the effects of galectin-3 (GAL-3) on the viability, migration and inflammation of human umbilical vein endothelial cells (HUVECs) and the mechanisms.METHODS:The HUVECs were cultured in vitro and treated with GAL-3 recombinant protein at 2 mg/L or GAL-3 short hairpin RNA (shRNA).The HUVECs were divided into normal group, recombinant GAL-3 group, shControl group and GAL-3-shRNA group.The mRNA expression of GAL-3, monocyte chemotactic protein (MCP)-1, IL-6, matrix metalloproteinase (MMP)-9 and cyclin D1 was detected by real-time quantitative PCR,and the protein expression of GAL-3, IL-6 and MCP-1 was detected by Western blot.The secretion levels of MCP-1 and IL-6 in the culture medium were measured by ELISA.The viability and the ability of migration of the HUVECs were examined by CCK-8 assay and wound healing assay.The protein levels of heat shock protein 90 (HSP90), ERK1/2, p-ERK1/2, JNK and p-JNK were determined by Western blot.RESULTS:The expression of GAL-3, MCP-1 and IL-6 at mRNA and protein levels, the mRNA expression of MMP-9 and cyclin D1, and the secretion levels of MCP-1 and IL-6 in the culture medium were significantly higher than those in normal group (P<0.05) after the HUVECs were treated with GAL-3 recombinant protein.However, these molecules mentioned above in GAL-3-shRNA group were significantly lower than those in normal group and negative control group (P<0.05).Compared with normal group, the viability and migration ability of the HUVECs in recombinant GAL-3 group were significantly increased, but the viability and migration ability of the HUVECs in GAL-3-shRNA group were lower than those in normal group and shControl group (P<0.05).In addition, the protein levels of p-ERK1/2 and HSP90 in recombinant GAL-3 group were higher than those in normal group (P<0.05), but those in GAL-3-shRNA group were lower than those in normal group and shControl group (P<0.05).The protein level of p-JNK was not oviously changed among the 4 groups.CONCLUSION:GAL-3 is involved in regulating the cell growth, migration and the release of inflammatory cytokines in vascular endothelial cells, which may be mediated by HSP90-ERK1/2 signaling pathway.

Objective To investigate the clinical efficacy of TCM external treatment combined with Dasangju Mixture in treating mycoplasma pneumoniae pneumonia (MPP) in children. Methods One hundred patients with MPP were randomly divided into control group and treatment group by parallel design, with 50 patients in each group. Patients in control group orally took azithromycin combined with other symptomatic treatment when necessary, 7 d for a course and continuously for 3 courses. Based on the treatment for control group, patients in treatment group were added with TCM external treatment (BL13 and RN17) combined with Dasangju Mixture, while azithromycin was sequentially used just for 2 courses. Other types of treatment lasted for 3 courses. The improvement time of symptoms and signs of the patients were recorded and the therapeutic effects of the two groups were evaluated. Moreover, the serum concentrations of mycoplasma pneumoniae-immune gloulin M (MP-IgM) were determined before and after treatment on the 1st and 21st days. Results The improvent time of fever, and cough and lung sign of the treatment group were more obviously shortened than the control group (P<0.05). The total effective rate was 96.0% (48/50) in observation group and 74.0% (37/50) in control group, with statistical significant difference between the two groups (P<0.05). After treatment, the negative rate of the serum concentration of MP-IgM in the treatment group was 82%, and that of the control group was 58%, with statistical significance between the two groups (P<0.05).Conclusion TCM external treatment combined with Dasangju Mixture can not only significantly improve the clinical symptoms and signs of MPP children patients, but also shorten the disease course, and the efficacy was significant. Meanwhile it can reduce the course and side effects of azithromycin, which provides reference to the administration of antibiotics in children.

Objective To compare differences and similarities of the content of trigonelline in Himalaica mirabilis of either wild or cultivated materials from different places. Methods An HPLC method was established to determine the content of trigonelline in Himalaica mirabilis. ZORBAX XDB-CN column (4.6 mm×250 mm, 5 μm) was used, with mobile phase of acetonitrile-0.03%acetic acid (85∶15), flow rate of 0.8 mL/min, detection wavelength of 265 nm, determination wavelength of 360 nm, and column temperature of 30 ℃. Results Regression calculation was made on peak area with the reference solution concentration, and then got the regression equation A=23.409C-26.398, r=0.999 8. Trigonelline showed good linear relation with peak area among the range of 2.004-200.400 μg/mL. The average recovery of trigonelline was 99.57%, RSD=1.11%. Conclusion There was no significant difference in the content of trigonelline of either wild or cultivated materials from different places. This study laid the foundation of application of the cultivated Himalaica mirabilis.

Objective To measure the expressions of programmed death?1(PD?1)and programmed death ligand?1(PD?L1)on peripheral blood T lymphocytes of patients with condyloma acuminatum(CA), and to investigate their role in cellular immunity in these patients. Methods Peripheral blood samples were obtained from 30 patients with CA(CA group)and 20 healthy human controls (control group). Flow cytometry was conducted to detect the expressions of PD?1 and PD?L1 on the surfaces of peripheral blood CD4+and CD8+T lymphocytes, and to determine the counts of CD4+and CD8+T lymphocytes. Enzyme?linked immunosorbent assay(ELISA)was performed to measure the levels of serum interleukin?2(IL?2)and interferon?γ(IFN?γ). Statistical analyses were carried out to compare the above parameters between the two groups, and to assess the relationship of PD?1 and PD?L1 expressions with the counts of CD4+and CD8+T lymphocytes as well as with the serum levels of IL?2 and IFN?γ. Results There was a significant increase in the expression rates of PD?1 and PD?L1 on CD4+T lymphocytes(PD?1:9.48%± 3.31%vs. 7.12%± 2.16%, t=2.81, P<0.01;PD?L1:4.40%± 1.46%vs. 3.26%± 1.13%, t=3.16, P<0.01)and CD8+T lymphocytes(PD?1:12.52%± 3.17%vs. 9.95%± 2.17%, t=3.16, P<0.01;PD?L1:7.07%± 2.23%vs. 5.39%± 1.69%, t=2.88, P<0.01)in the CA group compared with the control group. Moreover, the CA group showed significantly lower counts of CD4+T lymphocytes(727.43 ± 138.59/μl vs. 804.25 ± 92.83/μl, t=2.17, P<0.05)and CD4/CD8 ratio(1.23±0.35 vs. 1.46 ± 0.34, t = 2.24, P < 0.05) than the control group, while no significant difference was observed in CD8 + T lymphocyte counts between the CA group and control group(613.60 ± 121.60/μl vs. 572.45 ± 103.08/μl, t=1.24, P>0.05). The levels of serum IL?2 and IFN?γwere both lower in the CA group than in the control group(t=2.12, 2.16, respectively, both P < 0.05). In the CA group, PD?1 and PD?L1 expression levels on peripheral blood CD4 + T lymphocytes were both negatively correlated with CD4+T lymphocyte counts, the CD4/CD8 ratio, as well as IL?2 and IFN?γserum levels(all P<0.05), and those on peripheral blood CD8+T lymphocytes were also negatively correlated with the CD4/CD8 ratio(all P<0.05), but uncorrelated with CD8+T lymphocyte counts(both P>0.05). Conclusion PD?1 was highly expressed on peripheral blood T lymphocytes from patients with CA, which may inhibit T lymphocyte?mediated immune response, decrease CD4+T lymphocyte counts, the CD4/CD8 ratio as well as IL?2 and IFN?γserum levels by interacting with its ligand PD?L1 and forming the PD?1/PD?L1 signaling pathway.

This study was aimed to establish the TLC identification method of Radix Mirab ilis himalaic a. The β-sitosterol and daucosterol were used as the reference substances. The single-factor test was used. A variety of factors which affected TLC were systematically investigated to filter out the best TLC conditions for identification of different batches of medicines. The results showed that the best TLC conditions were as follows: silica gel G plates, extraction solvent (methanol), reagent (5% sulfuric acid in ethanol), extraction method (ultrasonic extraction with methanol), ex-tracted time (30 min), the agent (petroleum ether-ethyl acetate-acetone (5:2:1)) and sample volume (6 μL). It was concluded that the method, which had high separation degree, was reproducible and simple. It can be used as the quality control of Radix Mirab ilis himalaic a.

Objective To explore the application of optical coherence tomography in vascular tissue engineering culture by dynamic monitoring its changes.Methods Human umbilical artery smooth muscle cells were isolated and culture by tissue block method.After passage culture and cell surface markers evaluation, smooth muscle cells were seeded onto polyglycolic acid scaffold and placed into the bioreactor based on Luo-Ye pump with pulsatile stress for three-dimensional culture.At 1、4、 7 、10、14、17、21 days in culture, the image data was obtained by optical coherence tomography technology.The ability of imaging TEBV via OCT was analyzed combined with histopathological observation.Results As the incubation time extended,OCT clearly showed PGA gradual degradation, decreased composite scaffold thickness and the wall structure from loose to tight.At 21 days in culture, the vessel mimics had smooth surface with extracellular matrix evenly distributed and achieved complete reconstruction in the PGA scaffold.Combining with histopathological staining, the blood vessel mimics were similar to natural blood vessels.OCT measured TEBV thickness compared with histopathological measurement had good correlation (r =0.922,P ＜ 0.05).Conclusion Optical coherence tomography could clearly image microstructures of tissue engineered blood vessels cultured in three-dimensional culture system based on Luo-Ye pump, delineate the reconstruction of TEBV-like tissue in the bioreactor and provide as a dynamic and convenient monitoring tool in vascular tissue engineering.

Background Ocular alkali burns leads to corneal ulcer and angiogenesis and even corneal opacity.There is still no ideal treatment method.Studies showed that mesenchymal stem cells (MSCs) can repair corneal wound in vivo,but the specific mechanism is still not clear.Objective This study aimed to observe the histopathological change after the early transplatation of bone marrow MSCs (BMSCs) for corneal alkali burn model in rabbits and explore the anti-inflammatory effects of MSCs after corneal alkali burn.Methods Bone marrow of 4 ml was collected from 2-3 month-old Japanese rabbit.BMSCs were isolated and cultured from the bone marrow of rabbits,and the third generation of cells were used in this study.Cultured cells were identified by morphology and the expressions of surface markers.Corneal alkali burn models were extablished in the right eyes of 24 rabbits by attaching the filter paper with 0.1％ NaOH at the central cornea for 30 seconds,and then the models were randomized into 2 groups.BMSCs suspension of 300 μl (concentration 5×l06/μl) was subconjunctivally injected 1 hour after modeling in the BMSCs group,and equal volume of PBS was used in the same way in the PBS group.Corneal opacification was scored under the slim lamp microscope in 3,14 and 28 days after injection.The polymorphonuclear neutrophils (PMNs) were counted by histopathological examination,and the expression of matrix metalloproteinase-2 (MMP-2) in the corneal tissue was evaluated by immunochemistry in various time points.The use and care of the rabbits followed the statement of ARVO.Results The rabbit BMSCs were plastic-adherent cells that exhibited a fibroblastlike shape.Cultrued cells highly expressed surface adhesion molecular markers CD29 and CD90 (99.18％ and 97.94％) and lowly expressed hematopoietic cell markers CD34 and CD31 (0.74％ and 0.15％).Opacification of cornea,defect of corneal epithelium,stromal edema and neovascularization appeared after modeling.In 14 days and 28 days after modeling,the opacification scores in the BMSCs group were 2.37±0.52 and 2.25±0.50,which were significantly lower than 3.00±0.53 and 3.25 ±0.50 in the PBS group (t =2.376,2.828,both at P＜0.05).After subconjunctival injection,the number of PMNs was (34.17 ±1.85) /12 fields and (25.64 ±3.86)/12 fields in the BMSCs group,showing significant decrease in comparison with (42.70 ±1.54) /12 fields and (32.67 ±1.42)/12 fields in the PBS group (t=10.021,4.832,both at P=0.000).The expression levels of MMP-2 (A value) in cornea were 0.388±0.016 and 0.384±0.006 in the BMSCs group,with considerable decreases in comparison with 0.438± 0.006 and 0.412± 0.005 in the PBS group (t=10.205,13.514,both at P=0.000).Conclusions Early transplantation of BMSCs can arrest the occurrance of corneal ulcer by suppressing the infiltration of PMNs,alleviateing the inflammation reaction,downregulating the expression of MMP-2 in cornea and inhibiting the degradation of stromal collagen fibers.

Objective Using literature metrology,scientific literature in the field of depression models in animals was analyzed to better understand the development trends and hot topics in this field.Methods We obtained publications on depression and animal models from 2013 to 2022 from the Web of Science core set database.CiteSpace 6.1 R1(64-bit)Basic was used to analyze annual publications,countries,institutions,authors,and keywords related to this field.Results A total of 1000 articles were included in this study.From 2013 to 2022,the number of articles published increased gradually and then stabilized.In terms of the number of articles,the United States had the most published articles(256).Wegener Gregers was the most influential author in the field with 23 published articles.Conclusions The field focuses on signaling pathways and therapeutic approaches to determine the pathogenesis of depression and better treatments.This study provides a visual analysis of trends in depression research to help researchers keep up with the latest developments.

Objective The present situation and hotspots of KM mouse research were visualized by CiteSpace software,which provided a reference for breaking through the bottleneck of KM mouse research and finding new ideas in China.Methods A total of 3981 articles were retrieved from CNKI and Web of Science databases from January 1,2004 to June 10,2023,of which the number of publications,countries(regions),institutions,author cooperation network,keyword co-occurrence,clustering,burst words,and cited English literature for analysis.Results The number of documents published in Chinese was decreased and the number of documents published in English was increased gradually in KM mouse research.The three countries with the most published articles were China,the United States,and Japan,where the leading research institutions were the China Agricultural University,Chinese Academy of Sciences,and Nanjing University of Chinese Medicine.The main authors at home and abroad were Liu Weihui,Li Guangwu,and Zhao Xin.The domestic research focused on the toxicology,immunology,and reproduction of KM mouse,and the foreign research focused on the cell biology and physiology of KM mouse.Conclusions KM mouse have been used and popularized in our country for more than 70 years,mainly focusing on toxicology,immunology,and reproduction.Although the use of KM mouse has led tove made allowed some achievements in experiments,certain it is necessary to solve their problems in the future,such as the standardization of population breeding,the discovery of the dominant research fields,and the further promotion of animal ethics,should be resolved.