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
;
Biomarkers
;
blood
;
Blood Proteins
;
metabolism
;
Humans
;
Male
;
Middle Aged
;
Silicosis
;
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.
6.Foot hyalohyphomycosis caused by Fusarium subglutinans:a case report
Qiu-Xia CHEN ; Wen-Ming HUANG ; Jian-Qiang SHI ; Wen LI ; Shun-Fan LI ; Rong-Xi LU ; Feng-Yan BAI
Chinese Journal of Dermatology 2003;0(08):-
Objective To report a case of foot hyalohyphomycosis due to Fusarium subglutinans. Methods Medical history was collected and physical examination performed for this patient.Biopsy samples were obtained from the inner side of right ankle of this patient and subjected to pathological examination. Discharge was collected from the lesions for direct microscopic examination and culture.Results A 72-year-old woman presented with an ulcer on the right foot for 3 years.Physical examination disclosed an ulcer,measuring 3 cm x 1.5 cm,with a moist surface and obvious tenderness,at the inner side of the right ankle.Proliferation of dusky-red granulomatous tissue was observed at the base of the ulcer.Pathological examination revealed necrotic granulomatous tissue and slender,septate and hyaline hypha-like structure in the superficial dermis with scattered infiltration of inflammatory cells.PAS staining showed sausage-like hypha and scattered orbicular-ovate spores.Microscopic examination of lesional discharge exhibited septate, branching and hyaline hypha.The isolated fungus was identified as Fusarium subglutinans by culture,and appeared to be highly sensitive to terbinafine,nystatin and amphotericin B.The lesion completely healed after 2 months of treatment with oral terbinafine (0.25 g,twice a day).Conclusions This is a case of foot hyalohyphomycosis due to Fusarium subglutinans,and terbinafine is effective for this condition.
8.Effect of Electroacupuncture at the Beginning and Ending Points of Bicipital Muscle on the Superficial Electromyography of the Spastic Limb in Hemiplegia
Tingting YAN ; Xiang LI ; Hongrui ZHANG ; Huanqing FAN ; Wen ZHANG ; Huan GUO ; Zili XU
Shanghai Journal of Acupuncture and Moxibustion 2017;36(4):379-382
Objective To observe the effect of electroacupuncture at the beginning and ending points of bicipital muscle on the superficial electromyography (sEMG) of the spastic limb in hemiplegia.Method Forty patients with spastic hemiplegia after cerebral stroke were divided by the random number table into a group of electroacupuncture at the beginning and ending points (group A) and a group of electroacupuncture at antagonistic muscles (group B). The former selected Ashi points at the beginning and ending points of bicipital muscle, while the latter selected points at the antagonistic muscles [Tianjing (TE10), Qinglingyuan (TE11), etc.], to receive perpendicular puncturing. The needles were retained for 30 min, and the sEMG of bicipital muscles in resting state was detected after the removal of the needles. The spastic bicipital muscle was examined by sEMG prior to the acupuncture treatment, and respectively after 2-week and 4-week acupuncture treatment, and the detected parameters included root mean square (RMS) and integrated electromyography (IEMG).ResultThe EMG and IEMG declined gradually in the two groups after the acupuncture treatment; the intra-group comparisons of the RMS and IEMG values at three time points, e.g. prior to acupuncture treatment, after 2-week acupuncture treatment and after 4-weekacupuncture treatment, showed that the values changed significantly compared to those at the previous time point (P<0.05); the between-group comparisons showed that there were no significant differences in comparing the RMS and IEMG values regardless of different time points (P>0.05). The RMS and IEMG values presented same changing tendencies after 4-week acupuncture treatment in the two groups.ConclusionAcupuncture at the beginning and ending points and at the points on antagonistic muscles both can decrease the resting-state muscle tension in hemiplegia patients; sEMG is of certain significance in evaluating the treatment of hemiplegia.
9.Effect of rosiglitazone on expression and excretion of connective tissue growth factor in renal tissue and urinary of diabetic rats
Wei CHEN ; Shandong YE ; Yingxin WANG ; Wen HU ; Aihong FAN ; Yan CHEN ; Guangwei YANG ; Xiucai LI
Chinese Pharmacological Bulletin 2003;0(09):-
Aim To study the effect of rosiglitazone(RGZ)on expression and excretion of connective tissue growth factor(CTGF) in renal tissue and urinary of diabetic nephropathy rats.Methods Three groups of rats were studied:normal control group(n=8),STZ-induced diabetic model group(n=8),diabetes RGZ-treatment group(n=8).Urinary CTGF was measured at the 1 st,4 th,6 th and 8 th week,as well as the expressions of CTGF protein(by histochemical staining)at the 8 th week by ELISA.Results The urinary excretion rates of CTGF at 4 th、6 th、8 th weeks and the expressions of CTGF protein in renal cortex at 8th weeks significantly increased in STZ-induced model group compared with those in normal group(P
10.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
;
Biomarkers
;
blood
;
Chromatography, High Pressure Liquid
;
Diabetes Mellitus, Experimental
;
blood
;
Diabetes Mellitus, Type 2
;
blood
;
Metabolome
;
Phospholipids
;
blood
;
Rats
;
Rats, Sprague-Dawley
;
Tandem Mass Spectrometry