1.Construction of Recombinant Adenovirus Expression Vector of Human Sema4C Gene and Its Expression in Mouse Myoblasts Cell Line C2C12
Hai-Tao WU ; Shu-Hong LIU ; Yan WU ; Jun-Die FAN ; Wen-Hong FAN ; Ming FAN ;
China Biotechnology 2006;0(04):-
To generate recombinant adenovirus expression vector of human Sema4C gene and observe its expression in mouse myoblasts cell line C2C12 for ensuring easy access to investigate the role of Sema4C gene during myogenesis. The recombinant plasmid was packaged and amplified after being transfected in HEK293 cells through Lipofectamine. After infecting C2C12 myoblasts with recombinant adenovirus vector, the adenoviral infection efficiency was determined by confocal microscope which showed that the expression of green fluorescence could be detected at 12h and then reached peak at 24h after recombinant adenovirus infection. The infection efficiency was almost 100% confirmed by FACS examination. Detection of WB indicated that the expression of Sema4C in C2C12 of recombinant adenoviral infection group was significantly higher than that of the control group (P
2.Expression of bmp3,bmp4 and bmp7 during dog permanent tooth root formation
Kun XUAN ; Fusheng YANG ; Lingying WEN ; Yan JIN ; Shumei FAN
Journal of Practical Stomatology 2000;0(06):-
Objective:To evaluate the physiological roles of bmp3,bmp4 and bmp7 during the formation of permanent tooth roots in dog. Methods:The expression of bmp3,bmp4 and bmp7 mRNA at different stages of the development of permanent tooth roots was examined by in situ hybridization in 3 dogs aged 12-18 weeks. Results:bmp3 was found in dental sac surrounding the germs at the early stage of tooth root development, and in cementoblasts and periodontal cells at the later stage. bmp4 was found in odontoblasts, dental papilla and osteoblasts. bmp7 positive signals was found only in epithelial cells of root sheath around cervical circulus at early stage, then located in cementoblasts and odontoblasts at later stage. Conclusion:The spatiotemporal expressions of bmp3,bmp4 and bmp7 are widely diverse, indicating that they participate in the regulation of tooth root development.
3.Changes of plasma prethrombotic state molecular markers levels in silicosis patients.
De-Zhong ZHANG ; Jian-Yan WEN ; Li-Min FAN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2009;27(7):444-446
Adult
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Biomarkers
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blood
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Blood Proteins
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metabolism
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Humans
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Male
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Middle Aged
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Silicosis
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blood
4.Effect of perioperative intravenous flurbiprofen axetil on chronic postmastectomy pain
Mingyang SUN ; Qin LIAO ; Lingling WEN ; Xuebin YAN ; Fan ZHANG ; Wen OUYANG
Journal of Central South University(Medical Sciences) 2013;38(7):653-660
Objective:To explore whether perioperative intravenous flurbiprofen axetil can reduce the incidence and intensity of chronic pain for breast cancer atfer surgical treatment. Methods:This randomized, double-blind, controlled trial enrolled 60 patients undergoing mastectomy and axillary lymph node dissection under general anesthesia. All patients accepted Hospital Anxiety and Depression Scale (HAD) tests the day before the surgery to evaluate depression and anxiety. hTe patients were randomly assigned to receive either 50 mg lfurbiprofen axetil intravenously 15 minutes before the surgical incision and 6 hours later (group F) or intravenous 5 mL intralipid as a control (group C). All patients received patient-controlled intravenous analgesia (PCIA) with fentanyl postoperatively. Peripheral venous blood samples were drawn before the surgery, at 4 and 24 h atfer the surgery to detect the plasma level of PGE2 and tumor necrosis factor-α(TNF-α). Postoperative fentanyl consumption, Numerical Rating Scale (NRS) scores and adverse effects were recorded at 2, 6, 12, 24 and 48 h after the surgery. hTe duration and intensity of pain were followed up by telephone at the 2nd-12th month atfer the surgery. Results:The incidence of pain at 2, 4, 6, and 12 months after the breast surgery was 33%, 20%, 15%, and 10%, respectively, and the average pain score was 0.77, 0.57, 0.28, and 0.18, respectively. Compared with group C, the scores of pain in group F were significantly lower at 2, 4, 6 and 12 months postoperatively (F=7.758, P=0.007). The incidence of pain in group F was significantly lower at 2, 4 and 6 months postoperatively (P<0.05). There was no significant difference in the incidence of pain between the groups at 12 months postoperatively (P>0.05). Preoperatively and at 4 and 24 h atfer the surgery, there was no signiifcant difference in the level of TNF-αbetween the two groups (F=0.530, P=0.470);but plasma concentration of PGE2 in group F was significantly lower than that in group C (F=5.646, P=0.021). No patients developed abnormal bleeding, peptic ulcer, impaired liver or renal function and respiratory depression. Conclusion:Perioperative intravenous infusion of 100 mg flurbiprofen axetil can decrease the intensity and incidence of chronic pain for breast cancer atfer surgical treatment.
5.Anti-SARS-CoV-2 activity of small molecule inhibitors of cathepsin L
Wen-wen ZHOU ; Bao-qing YOU ; Yi-fan ZHENG ; Shu-yi SI ; Yan LI ; Jing ZHANG
Acta Pharmaceutica Sinica 2024;59(3):600-607
The coronavirus disease 2019 (COVID-19) is an acute infectious disease caused by the new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, which has led to serious worldwide economic burden. Due to the continuous emergence of variants, vaccines and monoclonal antibodies are only partial effective against infections caused by distinct strains of SARS-CoV-2. Therefore, it is still of great importance to call for the development of broad-spectrum and effective small molecule drugs to combat both current and future outbreaks triggered by SARS-CoV-2. Cathepsin L (CatL) cleaves the spike glycoprotein (S) of SARS-CoV-2, playing an indispensable role in enhancing virus entry into host cells. Therefore CatL is one of the ideal targets for the development of pan-coronavirus inhibitor-based drugs. In this study, a CatL enzyme inhibitor screening model was established based on fluorescein labeled substrate. Two CatL inhibitors IMB 6290 and IMB 8014 with low cytotoxicity were obtained through high-throughput screening, the half inhibition concentrations (IC50) of which were 11.53 ± 0.68 and 1.56 ± 1.10 μmol·L-1, respectively. SDS-PAGE and cell-cell fusion experiments confirmed that the compounds inhibited the hydrolysis of S protein by CatL in a concentration-dependent manner. Surface plasmon resonance (SPR) detection showed that both compounds exhibited moderate binding affinity with CatL. Molecular docking revealed the binding mode between the compound and the CatL active pocket. The pseudovirus experiment further confirmed the inhibitory effects of IMB 8014 on the S protein mediated entry process.
7.UPLC-Q-TOF/MS analysis of phospholipids metabolite profiling in plasma of type 2 diabetes mellitus rat.
Ping-yan ZENG ; Yu ZHANG ; Wen RUI ; Xia WU ; Yi-fan FENG
Acta Pharmaceutica Sinica 2015;50(7):882-886
This study reported the analysis of plasma phospholipid metabolism of the rats and the pathological biomarkers between the type 2 diabetes model control group (MC) and the normal control group (NC). SD rats were randomly divided into 2 groups: NC and MC. To investigate state of plasma metabolite profiling in normal body, type 2 diabetes mellitus (T2DM) model group using UPLC-Q-TOF/MS which was used as analysis tool in this research. The compounds were identified by UPLC-Q-TOF/MS based on MS/MS fragment ions information, element composition in MassLynx 4.1 and the Lipid Maps database. The sign of two groups of samples in specific markers for screening was through a software package in R software (BioMark software). The results show that the pathological markers were mainly phosphatidylcholine (PC) and triglycerides (TG); the 2-acyl PC in the MC group was less more obviously than that in the NC group; high carbon number and high degree of unsaturation of the TG was reduced under the condition of type 2 diabetes. In the state of type 2 diabetes, metabolic changes occurred in rat plasma phospholipids obviously, which had a close relationship with the occurrence and development of T2DM.
Animals
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Biomarkers
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blood
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Chromatography, High Pressure Liquid
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Diabetes Mellitus, Experimental
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blood
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Diabetes Mellitus, Type 2
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blood
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Metabolome
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Phospholipids
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blood
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Rats
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Rats, Sprague-Dawley
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Tandem Mass Spectrometry
8.Application of milk duct scope in bilateral nipple discharge patients with hypothyroidism or pituitary tumor
Meixiang LUAN ; Jiarong FAN ; Hao WEN ; Shuyi YANG ; Yan FU ; Lati MU ; Li LIU
Chinese Journal of General Surgery 2009;24(9):725-727
Objective To investigate whether the bilateral nipple discharge with the background of hypothyroidism and pituitary tumor causes intraductal neoplasm. Methods Clinical data of 222 bilateral nipple discharge patients with hypothyroidism or pituitary tumor undergoing mammary ductoseopy (MDS) examination were analyzed. Those diagnosed as intraductal papilloma received surgery and pathological exam. 158 cases with inflammatory comedomastitis diagnosed by MDS received milk duct washing with gentamycin, dexamethasone and corresponding medical treatment. Results Among the 222 cases, 158 cases(71.17%) were diagnosed as inflammatory comedomastitis by MDS, 64 cases (28.83%) were diagnosed as intraduetal papilloma (64/222) by MDS and received operation,59 cases were pathologically diagnosed as intraductal papilloma,the other 5 cases were pathologically diagnosed as mammary duct ectasia. The pathological coincidence was 92.2% and no breast cancer was observed. 68.10% of inflammatory comedomastitis cases suffered from breast pain. There were no significant difference in the disease distribution of the ethnic groups. During pre-menopause period, the incidence of period neoplasia is higher in pituitary tumor than hypothyroidism; However, during the post-menopause the situation was vice versa. The color of niplle discharge is mostly milk-like in patients with pituitary tumor, while serous in patients with hypothyroidism. 71.19% of duct papilloma locate at the branch of the main duct. Conclusion The bilateral nipple discharge with hypothyroidism or pituitary are usually concomitant with intraduetal lesions. Mammary ductosopy is helpful to discover intraduetal neoplasia.
10.Study on bone marrow mesenchymal stem cells in repairing of radiation induced acute liver injury of rats
Yongxing BAO ; Fan LOU ; Huarong ZHAO ; Huhu ZHU ; Yan MA ; Hao WEN
Chinese Journal of Radiological Medicine and Protection 2010;30(1):22-25
Objective To investigate the role of mesenchymal stem cells in the repair of radiation induced liver injury.Methods 12 female SD rats were irradiated with 20 Gy 6 MV X-rays on the right lobe of the liver,to establish the model of radiation induced liver injury.The rats were divided randomly into two groups as invention group and control group,and transplanted with 1 ml male mesenchymal suspension or 1 ml normal saline in 4 hours after radiotherapy.The morphological changes of liver were observed.The existence of sex determining gene Y(SRY)and the level of alpha-smooth muscle actin (a-SMA) were detected.Results Some injury of right lobe liver in two groups were observed,and the injury degree of right lobe liver in intervention group were lower than that of control group.The amount of SRY positive cells in the right lobe liver of intervention group was higher than that in the left lobe liver(t = 3.77,P <0.05).The positive expression rate of a-SMA in right lobe liver of intervention group was lower than that of control group.Conclusions Acute radiation induced liver injury could lead BMSCs' homing in order to decrease the degree of liver fibrosis.