Objective:To investigate the effects of Penehyclidinehydrochloride(PHCD) on endotoxin-induced cerebral edema.Methods:one handred and five S-D male rats(200～220g)were randomly divided into 5 groups(n=21):groupC control;groupL LPS;group DL,DM and DH received intraperitoneal PHCD 0.05,0.15 or 0.45mg/kg 10min before lipopolysaccharide(LPS) administration.Cerebral edema was induced by internal carotid arterial LPS 150?g.Seven animals each group were decapitated at 4、6 and 12h after operation and their brains were immediately removed for determination of water content of brain and microscopic examination.Results:Administration of LPS caused severe brain damage and significantly increased water content of the brain.The LPS-induced changes were mitigated by pretreatment with different doses of PHCD in groupDM and DH,but there was no significant difference between the two groups.Conclusion:Pretreatment with PHCD can attenuate LPS-induced cerebral edema.

Objective To investigate the effects of penehyclidine hydrochloride (PHCD) pretreatment on expression of NF-κB and iNOS in rats with LPS-induced brain injury. Methods One hundred and five male SD rats weighing 200-220 g were randomly divided into 5 groups (n=21 each): group Ⅰ normal saline (NS);group Ⅱ LPS (L);group Ⅲ,Ⅳ,Ⅴ PHCD 0.05, 0.15, 0.45 mg/kg (D1,2,3). The animals were anesthetized with chloral hydrate 350 mg/kg. Brain injury was induced by intra-arterial LPS 150 μg administered via left internal carotid artery in group Ⅱ-Ⅴ. In group Ⅲ,Ⅳ, and Ⅴ PHCD 0.05, 0.15 and 0.45 mg/kg were given intraperitoneally (IP) at 10 min before intra-arterial LPS. The animals were decapitated at 4, 6 and 12 h after administration of PHCD (n=7 at each time point in each group). The brains were immediately removed for determination of water content, expression of NF-κB and iNOS protein and examination with light and electron microscope. Results Water content of the brain and expression of NF-κB and iNOS protein were significantly higher in group L, D1, D2 and D3 than in group NS and were significantly lower in group D2 and D3 than in group L. Intra-arterial LPS produced severe damage to the brain which was significantly attenuated by PHCD in group D2 and D3. Conclusion PHCD 0.15,0.45 mg/kg pretreatment can attenuate LPS-induced brain injury by inhibiting the up-regulation of expression of NF-κB and iNOS.

Objective To analyze the multi-slice computed tomography (MSCT) enhanced findings in the patients with primary small intestinal stromal tumor(SIST),and to probe the relationship between the imaging findings and the pathologic risk in order to improve the diagnostic accuracy.Methods Thirty patients with primary SIST confirmed by surgical pathology were enrolled in this study.Characterization and compassion of the clinical manifestations and MSCT enhanced findings were carried out between the pathologic low-and high-risk groups.Furthermore,the relationship was analyzed between the enhanced findings and the pathologic risk.Results Among all 30 patients with primary SIST,the lesion was located at duodenum in 5 patients (16.7％),at jejunum in 16 (53.3％),and at ileum in 9 (30％).14 patients were classified in the low risk group with the lesion with the average length of (3.8±0.9) cm,and other 16 in the high-risk group with lesion with the average length of (7.0 ± 1.4) cm.There were no statistical differences between the low-and high-risk groups in CT value in plain and venous phase,and in added value in arterial,venous and delayed phases.However,the significantly differences were observed in CT value in arterial and delayed phases between two groups (P＜0.05).Conclusion MSCT may effectively evaluate the pathologic risk of primary SIST.There are significant differences of the enhanced findings between the low and high-risk groups,which can provide important apreoperative classification for the therapy.

Objective To investigate the effect of propofol on the learning and memory function in neonatal rats under hypoxic conditions. Methods Eighty-four 7-day-old SD rats were randomly divided into 6 groups (n = 14 each): propofol + 18% oxygen (propofol-hypoxia, group PH), propofol + air (group PA), propofol +100% oxygen (propofol-oxygen, group PO), 0.9% normal saline (NS) + 18% oxygen (group CH), NS + air (group CA), NS + 100% oxygen (group CO). The rats received injection of intraperitoneal propofol 50 mg/kg or NS 5.0 ml/kg once a day for 7 consecutive days and they were exposed to 18% oxygen, air or 100% oxygen at the end of each injection. SaO2 and respiratory rate (RR) were monitored and recorded after administration. The rats were returned to the cage after recovery of the righting reflex. Six rats in each group were sacrificed 24 h after the 7th injection, and the brain tissues were taken to observe the apoptosis in hippocampal neurons. Morris water maze test was used to test the learning and memory function 2 weeks after administration in the other rats. Results RR was significantly lower and the escape latency at T1.2 longer in group PO than in group CO (P ＜ 0.05). RR and SaO2 were significantly decreased, apoptotic index was increased, the escape latency was prolonged and the frequency of crossing the original platform was reduced in group PA compared with group CA, and in group PH compared with group CH (P ＜ 0.05). Compared with group PO, SaO2 was significantly decreased, apoptotic index was increased, the escape latency was prolonged and the frequency of crosing the original platform was reduced in group PA (P ＜ 0.05). Conclusion Propofol induces apoptosis in hippocampal neurons and decreases the learning and memory function in neonatal rats under hypoxic conditions.

Objective:To observe the protective effect of Zhicao Tea Mixture on Müller cells and the expression of inflammatory factors in mice with diabetic retinopathy.Methods:Seventy-five C57BL/6J mice were randomly divided into the normal control group, diabetes mellitus (DM) group, low concentrations group, medium concentrations group and high concentrations group, with 16 mice in each group. The diabetes model of mice in all groups except the normal control group were established by intraperitoneal injection of STZ (60 mg/kg). Four weeks after the successful modeling, the Zhicao Tea Mixture with low (30 ml/kg), medium (60 ml/kg) and high concentrations (120 ml/kg) were respectively administered by gavage. Weight and blood glucose of mice in each group were measured every two weeks. After 8 weeks, Western blot method was used to detect the mice retina Müller cells activation marker gelatinous fibrous acidic protein (GFAP). Immunofluorescence was performed to detect the expression GFAP and glutamine synthetase (GS). Real-time quantitative PCR (RT- qPCR) and ELISA were used to determine the mRNA and protein expression levels of mouse retinal VEGF, TNF-α, IL-1β and IL-6 respectively.Results:The weight of mice in the DM group was lower than that of the normal control group, and the blood glucose was increased. Zhicao Tea Mixture had no effect on the weight of DM mice, but had a significant hypoglycemic effect. The GFAP expression ( t=38.318, P<0.001) in the retina of mice in the DM group was increased and GS expression ( t=29.737, P<0.001) was decreased compared with the control group. The GFAP expression ( t=13.677, 19.387, 16.305; P<0.05) in the retina of mice in the low, medium and high concentrations group were decreased and GS expression ( t=5.170, 19.399, 6.705; P<0.05) were increased compared with the DM group. The expressions of retinal inflammatory factors VEGF, TNF-α, IL-1β and IL-6 in DM group all increased, while the expressions of the above-mentioned inflammatory factors in the retina of mice decreased in the low, medium and high concentrations group. Conclusion:Zhicao Tea Mixture can decrease the blood glucose of DM mice and reduces the diabetic retinal inflammatory response.

CD146 is a newly identified endothelial biomarker that has been implicated in angiogenesis. Though in vitro angiogenic function of CD146 has been extensively reported, in vivo evidence is still lacking. To address this issue, we generated endothelial-specific CD146 knockout (CD146(EC-KO)) mice using the Tg(Tek-cre) system. Surprisingly, these mice did not exhibit any apparent morphological defects in the development of normal retinal vasculature. To evaluate the role of CD146 in pathological angiogenesis, a xenograft tumor model was used. We found that both tumor volume and vascular density were significantly lower in CD146(EC-KO) mice when compared to WT littermates. Additionally, the ability for sprouting, migration and tube formation in response to VEGF treatment was impaired in endothelial cells (ECs) of CD146(EC-KO) mice. Mechanistic studies further confirmed that VEGF-induced VEGFR-2 phosphorylation and AKT/p38 MAPKs/NF-κB activation were inhibited in these CD146-null ECs, which might present the underlying cause for the observed inhibition of tumor angiogenesis in CD146(EC-KO) mice. These results suggest that CD146 plays a redundant role in physiological angiogenic processes, but becomes essential during pathological angiogenesis as observed in tumorigenesis.
Animals ; CD146 Antigen ; genetics ; metabolism ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Female ; Fibrosarcoma ; metabolism ; pathology ; Male ; Melanoma, Experimental ; metabolism ; pathology ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; NF-kappa B ; metabolism ; Neovascularization, Physiologic ; drug effects ; Phosphorylation ; drug effects ; Proto-Oncogene Proteins c-akt ; metabolism ; Retinal Vein ; growth & development ; pathology ; Signal Transduction ; drug effects ; Transplantation, Homologous ; Vascular Endothelial Growth Factor A ; pharmacology ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

Objective : To examine the role of LMO4 in the regulation of endothelial cell differentiation and angio- genesis in murine embryonic stem cells (mESC) ． Methods : Mouse Lmo4 cDNA was obtained from MEL cells by using the reverse transcription-polymerase chain reaction (RT-PCR) and subcloned into the expression vector pFG to generate the pFLG ，in which contained Flk-1 promoter to drive Lmo4 expresses in only FLK-1 + cells．The mESC were transfected with pFG or pFLG plasmids and subsequently screened with geneticin ( G418) to produce cell clones． These cell clones were named mESC /pFG and mESC /pFLG ，respectively． The mESC /pFG and mESC /pFLG were cultured in the differentiation medium for either 4 days or 10 days to generate embryoid bodies (EB) ．The 10-day embryoid bodies ( 10 d-EBs) carrying the pFG and pFLG vectors were subsequently stimulated to generate the blast-colony forming cells (BL-CFC) ，which indicated the presence of hemangioblasts．The endo- thelial cell sprouting analysis was performed by using 10 d-EBs．The expression of the interest genes was detected by using qualitative RT-PCR or Western blot analysis. Results : The pFLG expression vector was successfully con- structed through PCR identification．The mESC /pFG and mESC /pFLG cells were obtained after transfected with the pFG or pFLG vectors and selected by G418．The cells spontaneously differentiate to generate EBs，in which some green fluoresce cells were present．Western blot analysis showed that a significant increase in LMO4 expression in both 4 d-EB and 10 d-EB when compared to mESC．BL-CFC analysis showed that the 4 d-EB/ pFLG had a higher cloning efficiency ( 7. 70% ± 1. 27% ) ，comparing with that of the 4 d-EB/ pFG ( 1. 15% ± 0. 48% ) ( P = 0. 021) ．Quantitative RT-PCR results showed that the expression of Flk-1，C-kit，Tie-2 and Ve-cad genes in 10 d- EBs /pFLG increased more than 2-fold compared to 10 d-EBs /pFG．The endothelial cell sprouting analysis result showed a significant increase in the number and length of new blood vessels in 10 d-EB/ pFLG compared to 10 d- EB/ pFG (P＜0. 05) ． Conclusion Overexpression of LMO4 promotes hemangioblast differentiation from mESC， and benefits for endothelial cell differentiation and angiogenesis.

Objective : To investigate lim domain protein 4 (LMO4) functions and mechanisms in regulating proliferation of skin squamous cells (A431) , the shRNAs targeted to human LMO4 were coated by calcium phosphate nanoparticles (NP) and transfected into A431 cells to inhibit LMO4 expression. Methods : Reverse transcription and quantitative polymerase chain reaction ( RT⁃qPCR) , immunohistochemistry analysis and Western blot were used to detect expression of the interest genes. The expression vectors with shRNA targeted to human LMO4 (NP/sh⁃L) were coated by the calcium phosphate nanoparticles , and transfected into A431 . The MTT assay was conducted to determine cell proliferation after transfected for 24 , 36 and 48 h. Cells were stained with propidium iodide and examined cell cycles by using flow cytometry. Results : LMO4 expressed at higher levels both in the skin squacalcium phosphate nanoparticles and DNA was 10 ∶ 1 . There was no significant difference of transfection efficiency between the NP/sh⁃L and lipofection approaches. The MTT assay showed that silencing LMO4 inhibited proliferadown did not alter expression of CDK4 and cyclin D1 . Conclusion The calcium phosphate nanoparticles could bind and transfer the foreign DNA into the targeted cells with high efficiency. Silencing LMO4 decreased expression of cyclin E and CDK2 resulted in inhibition of cell proliferation.

China has entered a stage of deep aging. The expanding aging population and subsequently an aging labor force pose significant challenges to China’s social and economic development. European Union (EU) countries have entered the aging phase earlier and to a greater extent. To address the labor shortage and the rising dependency ratio, the EU has implemented a series of policies and measures, including raising retirement age, promoting flexible retirement, enhancing work flexibility, improving welfare for elderly workers, creating age-friendly environments, focusing on occupational health, strengthening knowledge and skills training, and leveraging digital technologies in the workplace. These measures aim to prevent premature exit of elderly workers, better balance their work and life needs, improve occupational health and skills, and enhance overall work efficiency of companies. This review discussed the experience and lessons learned from the EU countries in addressing the aging workforce from the perspectives of occupational health and occupational skills development, aiming to provide rational suggestions to assist China in better adapting the challenges of aging workforce after steadily and orderly advancing the gradual reform of the statutory retirement age, thereby improving the efficiency of the overall workforce and the stability of the labor market, driving the talent dividend of highly educated and skilled middle-aged and elderly employees, and ultimately promoting sustained economic and social development.

Objective: To establish a Tohoku hospital pediatrics-1 (THP-1) cell line with G protein-coupled recep- tor108 ( GPR108) deletion and explore its functions． Methods: According to the requirements of the clustered regularly interspaced short palindromic repeats ( CRISPR) / CRISPR-associated protein 9 ( Cas9 ) system ，two single guide RNAs (sgRNA1 and sgRNA2) paring to the flanking fragments of human GPR108 gene were designed and synthesized．The two oligonucleotides were inserted in the pL-CRISPR. EFS．GFP vector to generate the new recombinant vectors ( pL-CRISPR. EFS．GFP-sgRNA1 and pL-CRISPR. EFS．GFP-sgRNA2 ) ．The recombinant vectors and packaging plasmids (pMD2. G and psPAX2) ，were co-transfected into 293T cells to generate virus for infecting THP-1 cells．The GFP + cells were screened and isolated in 96-well culture plates by flow cytometry to obtain single-cell clones．PCR and Western blot were used to detect whether GPR108 was successfully knocked out in THP- 1 cells．Both GPR108 + / + and GPR108 －/ － THP-1 cells were treated with lipopolysaccharide (LPS) ．Interleukin 8 (IL-8) derived from the THP-1 cells，which were treated by LPS，was detected with Western blot and cytometric bead array ( CBA) analysis. Results: The recombinant lentiviral vector pL-CRISPR. EFS．GFP-sgRNA was successfully constructed and single-cell clone F9 was obtained by flow cytometric sorting after transfection of THP-1 cells．PCR and Western blot both confirmed that F9 was a GPR108 －/ － THP-1 single-cell clone． LPS stimulated GPR108 －/ － and GPR108 + / + THP-1 cells，both Western blot and CBA results showed a significant decrease in IL- 8 synthesis and secretion in GPR108 －/ － THP-1 cells． Conclusion The GPR108 －/ － THP-1 cell clone is success- fully obtained based on the CRISPR / Cas9 system．GPR108 deletion in THP-1 cells treated by LPS leads to a decrease of IL-8 expression and secretion．It lays the foundation for further research on the molecular mechanisms of GPR108 in the immune inflammatory response.