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.Treatment of Level 2 Hypertension by Diagnosis and Treatment Program of Integrative Medicine: a Multi-centre, Randomized Controlled Trial.
Xi CHEN ; Guang-shu CHENG ; Jun-ming FAN
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(7):801-805
OBJECTIVETo explore the therapeutic effect of diagnosis and treatment program of integrative medicine (IM) on level 2 hypertension in the young and middle-aged patients and their ambulatory blood pressure.
METHODSA randomized, placebo parallel and controlled, multi-center clinical trial was performed. Totally 199 young and middle-aged level 2 hypertension patients were randomly assigned to the treatment group (99 cases) and the control group (100 cases). All received combined hypotensive treatment program by taking Nifedipine Sustained Release Tablet and Hydrochlorothiazide as basic drugs. Patients in the treatment group additionally took Western medicine (WM) combined Jiangyabao serial drugs (0.31 g per tablet, 2 tablets each time, twice daily), while those in the control group additionally took WM combined simulative agents of Jiangyabao serial drugs (0.31 g per tablet, 2 tablets each time, twice daily). The treatment course was 8 weeks for all, and 24-week follow-ups performed. 24 h ambulatory blood pressure and casual blood pressure, and their efficacies were compared between the two groups, and safety assessed as well.
RESULTSCompared with before treatment in the same group, daytime and night casual blood pressure, as well as 24 h ambulatory blood pressure were all obviously improved in the two groups (P < 0.01). Average diastolic and systolic blood pressures at night decreased more in the treatment group than in the control group with statistical difference (P < 0.05). There was no statistical difference in total efficacies of daytime casual blood pressure or ambulatory blood pressure (P > 0.05).
CONCLUSIONJiangyabao serial drugs combined WM in treating young and middle-aged level 2 hypertension patients showed obvious effect in improving night blood pressure, especially for night diastolic blood pressure.
Blood Pressure ; Blood Pressure Monitoring, Ambulatory ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Hypertension ; diagnosis ; therapy ; Integrative Medicine ; Middle Aged ; Nifedipine
3.Survey of prevalence of iodine deficiency disorders and iodine nutrition in high-risk areas of Inner Mongolia in 2007 and 2008
Zhi-zhong, ZHANG ; Jun, LIU ; Jie, FAN ; Xian-ming, LIU
Chinese Journal of Endemiology 2011;30(4):420-423
Objective To investigate new cases of endemic cretinism in high-risk areas of Inner Mongolia, the prevalence of endemic goiter, the implementation of comprehensive measures, and to provide scientific basis for developing control strategies against iodine deficiency disorders (IDD). Methods In 2007 and 2008, a search for new cretin cases was conducted among children under the age of 10 in 11 historical serious epidemic cretinism prevalent areas of Songshan district, Keshiketeng qi, Kalaqin qi, Zhalaite qi, Zhuozi county,Liangcheng county, Qingshuihe county, Helin county, Dongsheng district, Zhungeer qi and Yinjinhuoluo qi. Three to 5 towns were selected in each qi(county, district) and 3 to 5 village primary schools were selected in each town.One hundred and twenty to 200 children aged 8 to 10 were selected in each school to inspect thyroid by B ultrasound and palpation and to test intelligence and urinary iodine. Two villages were selected in each town and 30 households were selected to determine urinary iodine of housewives and salt iodine. Results A total of 56 cases of suspected cases of endemic cretinism were found from the 11 counties but no case was confirmed. The goiter rate of children aged 8 to 10 was 5.2% (309/5922) by palpation and 4.3% (252/5922) by B-ultrasound. The median urinary iodine of children aged 8 to 10 was 241.5 μg/L, and urinary iodine was 100 to 300 μg/L, accounted 65.9%(3901/5920). Median urinary iodine of the 9 Qis(counties, districts) was in the range of 200 and 300μg/L.Higher than 5% of the 11 Qis(counties, districts) with urinary iodine lower than 50 μg/L was zero. Higher than 10% of the 11 Qis(counties, districts) with urinary iodine lower than 100 μg/L was 1. The median urinary iodine of housewives was 225.6 μg/L, and lower than 50 μg/L accounted for 2.2%(35/1597). Higher than 10% of the 11 Qis(counties, districts) with urinary iodine lower than 100 μg/L was 4. A total of 2109 households were investigated and 97.45%(2055/2109) of them ate iodized salt and 2.55%(54/2109) of them ate non-iodized salt. Conclusions In the 11 investigated counties, goiter rate of children has decreased to less than 10%, and no new cretin was found. It could be concluded that the fulfillment of prevention and control of IDD is effective. The iodine nutrition of children and women of childbearing age is in an adequate level. The coverage rate of iodized salt has maintained at a higher level, the sales network is sound, the salt price is reasonable and salt is easy to get and the children's intelligence is protected effectively. But, the health education about IDD is still weak, need to be reinforced.
4.Trigemino-cervical reflex in patients with Kennedy' s disease
Ming LU ; Juyang ZHENG ; Shuo ZHANG ; Jun ZHANG ; Dongsheng FAN
Chinese Journal of Neurology 2009;42(8):529-531
Objective To study changes of trigemino-cervical reflex (TCR) in patients with Kennedy's disease (KD). Methods The parameters of TCR were analyzed among patients with KD, amyotrophic lateral sclerosis and healthy controls. Results The parameters of ipsilateral P19, N31, A and contralateral P19, N31, A among patients with KD were (23.91±4.84), (35.45±4.76) ms, 1.24± 0.33 and (24.34±4.82), (36.20±4.91) ms, 1.19±0.25, respectively. Compared with the healthy controls((18.37±2.16), (28.50±1.56) ms, 1.90±0.43; (18. 72±2. 18), (29. 19±1.43) ms, 1.84 ± 0. 40), the difference in each parameter was significant (ipsilateral : t = 5.77, 8. 19, -6. 64; contralateral:5.05, 7.62, -7.77, all P<0.01). Conclusion The parameters of TCR were abnormal in KD patients, indicating that the trigeminal nerves and the bulbar may be involved in the disease.
6.Effect of Panax notoginseng saponins on human kidney fibroblast.
Ying WEI ; Jun-ming FAN ; Li-ping PAN
Chinese Journal of Integrated Traditional and Western Medicine 2002;22(1):47-49
OBJECTIVETo study the effect of Panax notoginseng saponins (PNS) on cell proliferation, type I collagen secretion and integrin beta 1 expression in human kidney fibroblast (KFB).
METHODSKFB were cultured and stimulated by PNS in vitro. The cell proliferation, type I collagen secretion and integrin beta 1 expression in KFB were measured by methyl thiazolyl tetrazolium (MTT), enzyme-linked immunoabsorbent assay (ELISA) and flowcytometry respectively.
RESULTSWithin its optimal concentration range and effective time range, PNS could obviously inhibit the proliferation, type I collagen secretion and integrin beta 1 expression (all P < 0.05) in human KFB.
CONCLUSIONPNS would possibly be an effective drug for renal interstitial fibrosis prevention and treatment.
Cells, Cultured ; Collagen Type I ; metabolism ; Fibroblasts ; drug effects ; Fibrosis ; prevention & control ; Humans ; Integrin beta1 ; metabolism ; Kidney ; cytology ; pathology ; Panax ; chemistry ; Saponins ; pharmacology
7.Effect of total saponins of Panax notoginseng on transdifferentiation of rats' tubular epithelial cell induced by IL-1alpha.
Mi WANG ; Jun-ming FAN ; Xin-ying LIU
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(8):722-725
OBJECTIVETo study whether total saponins of Panax notoginseng (PNS) can prevent renal interstitial fibrosis occurrence through blocking the IL-1alpha induced tubular epithelial cell and decrease the secretion of extracellular matrix (ECM).
METHODSNormal rats' tubular epithelial cells NRK52E were cultured in vitro, their morphological changes were observed by inverted phase contrast microscope and scanning electron microscope. Flow cytometry technique and immuno-histochemical method was used to detect the expression of alpha-smooth muscle actin (alpha-SMA), and ELISA was used to quantitatively detect the fibronectin (FN) in the supernatant
RESULTSIL-1alpha could induce the transdifferentiation of tubular epithelial myofibroblast, showing hypertrophy of cells elongated and fusiform-shaped, with significantly enhanced expression of alpha-SMA and increased secretion of FN (P<0.05). After adding PNS of different concentrations, the morphology of cells restored close to normal tubular epithelial cells, with the increase of alpha-SMA expression and FN secretion significantly inhibited (P<0.05) in dose-dependent manner (P<0.05). But addition of different dosages of PNS alone showed no effect on tubular cells.
CONCLUSIONIL-1alpha could induce the transdifferentiation of tubular epithelial myofibroblast, promote the deposition of ECM component FN. PNS could inhibit IL-1alpha induced the transdifferentiation of NRK52E and secretion of ECM, therefore, PNS could be taken as a new drug for prevention and treatment of renal interstitial fibrosis and terminal stage of renal diseases.
Actins ; analysis ; Animals ; Cell Differentiation ; drug effects ; Cell Line ; Epithelial Cells ; cytology ; Fibronectins ; analysis ; Fibrosis ; prevention & control ; Interleukin-1 ; pharmacology ; Kidney ; pathology ; Kidney Tubules ; cytology ; Panax ; chemistry ; Rats ; Saponins ; isolation & purification ; pharmacology
8.Effects of different concentrations of chloroprocaine on KCNQ2/Q3 channel currents in HEK2936 cells
Shiji QIN ; Jun LI ; Hong LI ; Ming LI ; Fan ZHANG ; Senming ZHAO ; Yanxin CHENG
Chinese Journal of Anesthesiology 2013;33(8):944-947
Objective To evaluate the effects of different concentrations of chloroprocaine on KCNQ2/Q3 channel currents in HEK2936 cells.Methods Human embryonic kidney (HEK293) cells served as an expression system.KCNQ2 and KCNQ3 cDNAs and green fluorescent protein were transfected into HEK293 cells by using lipofectamine.The KCNQ2/Q3 currents were recorded by using the whole-cell patch-clamp technique.Part Ⅰ The transfected HEK293 cells were randomly divided into 4 groups (n =11 each):control group,and 1,10 and 100 mmol/L chloroprocaine groups.The KCNQ2/Q3 channel currents produced by different concentrations of chloroprocaine were recorded under different holding potentials (-40,0 and 40 mV) and the action time was 1 min.Part Ⅱ The transfected HEK293 cells were randomly divided into 2 groups (n =5 each):control group and 10 mmol/L chloroprocaine.The KCNQ2/Q3 channel currents were recorded under different holding potentials (-80-30 mV)and the action time was 1 min.Different test potentials were normalized and fitted to Boltzmann function,and KCNQ2/Q3 channel Ⅰ-Ⅴ curve was then obtained.The activation and deactivation currents were both fitted to a single exponential function and the time constants for current activation and for current deactivation were calculated.Results Part Ⅰ When the holding potential was 40,0 and-40 mV,the suppression rate of KCNQ2/Q3 channel currents in HEK293 cells was higher in 1,10 and 100 mmol/L chloroprocaine groups than in control group (P <0.05 or 0.01).Part Ⅱ Compared with control group,the time constant for the current activation at 0 mV of holding potential was prolonged,the time constant for the current deactivation was shortened when the holding potential was-80 mV,and the half-activation voltage of KCNQ2/Q3 channels was increased,the activation curve shifted to the depolarized potentials,and KCNQ2/Q3 channel Ⅰ-Ⅴ curve slope was decreased in 10 mmol/L chloroprocaine group (P < 0.05).Conclusion Chloroprocaine concentration-dependently suppresses KCNQ2/Q3 channel currents in HEK2936 cells.The KCNQ2/Q3 channel is closed in advance due to KCNQ2/Q3 channel opening delay induced by chloroprocaine thus decreasing the activity of KCNQ2/Q3 channels.
9.Pilot study of identifying retinal nerve fiber layer defects with optical coherence tomography
Li-na, HUANG ; Ning, FAN ; Xiao-li, SHEN ; Hong-bo, CHENG ; Ming-ying, LAI ; Jun, ZHAO
Chinese Journal of Experimental Ophthalmology 2011;29(6):530-534
Background Fundus photography is a traditional method for detecting local retinal nerve fiber layer (RNFL) defects,but the evaluation of its result depends on the observer's experience.The spectral domain optical coherence tomography (OCT) exhibit the defects of RNFL very clearly.Objective This study was to evaluate the diagnosis value and correlation between topographic profiles of localized RNFL defects determined by spectral domain and time domain OCT with fundus photography.Methods Forty-one normal eyes of 41 subjects and 55 eyes of 55 glaucomatous patients with localized,wedge-shaped RNFL defects identified by two glaucoma specialists in fundus photographs were enrolled in the clinical study.The angular location and width of RNFL defects determined on the images of fundus photography,Cirrus HD-OCT and Stratus OCT were analyzed respectively using Pearson's correlation coefficient and linear regression analysis.This study followed the Helsinki declaration and was approved by Ethic Committee of Shenzhen Eye Hospital.Written informed consent was obtained from each individual before the clinical examination.Results Seventy-five RNFL defects were identified in 55 eyes by two glaucoma specialists unanimously with the defect position at superior-temporal and inferior-temporal quadrants.The sensitivity of Cirrus HD-OCT and Stratus OCT to determining RNFL defects were 88.0% and 69.3% respectively and their specificities were 92.7% and 97.6% respectively.The angular locations of RNFL defects by Cirrus HD-OCT and Stratus OCT were highly correlated with those by fundus photography(r=0.993,r=0.992,P<0.001);while the angular widths of RNFL defects by Cirrus HD-OCT and Stratus OCT were moderately correlated with those by fundus photography(r=0.420,r=0.432,P=0.019,P=0.002).No significant differences were found in the defect width of RNFL between Cirrus HD-OCT or Stratus OCT and fundus photography(Cirrus HD-OCT:P=0.114;Stratus OCT:P=0.074),and significant difference was found in that between Cirrus HD-OCT and Stratus OCT(P=0.002).Conclusion Spectral domain OCT and time domain OCT can localize RNFL defects with high sensitivity and specificity.The measure value of Cirrus HD-OCT and Stratus OCT for RNFL defects shows a good diagnostic agreement with fundus photography.
10.Immunoregulatory effects of bone marrow-derived mesenchymal stem cells from NZBWF1/J mice on T lymphocyte subsets balance
Hui LI ; Jun LIANG ; Ya-Yi HOU ; Le-Ming FAN ; Ling-Yun SUN ;
Chinese Journal of Rheumatology 2003;0(08):-
Objective To investigate the immunoregulatory effect and compare the different regula- tions of bone marrow mescnchymal stem cells(MSCs)derived from both lupus(NZBWF1/J)and normal(BALB/ c)mice on T lymphocytes in vitro.Methods MSCs from NZBWF1/J and BALB/c mice bone marrow were iso- lated and expanded,and identified by the surface phenotypes.CD3~+ T lymphocytes isolated by nylon wool columns were stimulated by phorbol myristate acetate(PMA)and co-cultured with or without the two strains of MSCs for 24 h.Intracellular eytokines of T cell,such as interferon(IFN)-?,interleukin(IL)-4,IL-12,IL-6, were analyzed by flow cytometry and quantification of transcription factors T-box expressed in T cells(T-bet) and GATA-binding protein 3(GATA-3)were detected by reverse transcriptase PCR(RT-PCR).T cell apop- tosis was assessed by flow cytometry using rhodamine123.Results The results showed that a decrease of CD3~+ T cell apoptosis was seen when NZBWF1/J MSCs or BALB/c MSCs were added to T cells stimulated by PMA(P<0.05),and an increase of TH2 cytokines by NZBWF1/J MSCs and TH1 eytokines by BALB/c MSCs were observed in the CD3~+ T cells eo-cuhured with MSCs(P<0.05).Conclusion It is suggested that the al- teration of T subsets caused by MSCs may interfere with the systemic lupus erythematosus(SLE)development and normal MSCs may be effective in the improvement of SLE.NZBWF1/J MSCs have defective immunoregula- tory function when compared with MSCs from healthy mouse strains.