3.Effects of murine cytomegalovirus infected mouse embryonic fibroblasts on the proliferation and activation of co-cultured regulatory T cells
Yanan LI ; Feng FANG ; Hong ZHEN ; Yufeng ZHOU ; Yongsui DONG
Chinese Journal of Infectious Diseases 2008;26(11):641-646
Objective To explore the effects of murinecytomegalovirus(MCMV)infected mouse embryonic fibroblasts(MEF)on the proliferation and activation of regulatory T cell in vitro. Methods A co-culture system of T cell and MCMV infected MEF(T-MEF MCMV)was established.The viral load of supernatant was determined by plaque assay.The proliferation of T cells was observed with cell counting.The proportion of CD4+ CD25+ cells was measured by flow cytometry.The levels of Foxp3 protein were measured by Western blot.Reverse transcription polymerase chain reaction(RT-PCR)assay was utilized tO determine whether MCMV infection of MEF influenced the level of transforming growth factor(TGF)-β mRNA.The level of TGF-β protein in supernatant was measured by double-antibody sandwich enzyme linked immunosorbent assay(ELISA).The difference between T-MEF MCMV group and control group was assessed by one-way ANOVA.Results When T cells were co-cultured with MCMV infected MEF for 1 day and 3 days,the viral load in supernatant decreased.But when co-culture lasted for 6 days,the antiviral effect obviously diminished,as the viral load[(5.58±0.67)×105 PFU/mL]of the experimental group showed no statistic difference with MEF MCMV control group[(6.05±0.34)×105PFU/mL].When co-cultured with MCMV infected MEF for 3 days,T cell increased from pre-culture level of[(2.02±0.05)×106/mL]to(2.25± 0.13)×106/mL(P<0.05).But when co-culture lasted for 6 days,the number of T eelI returned to (2.08±0.14)×106/mL,which had no statistic difference with that of co-culture for 3 days system. Both the expressions of Foxp3 protein and the proportion of CD4+ CD25+ Foxp3+ Treg cells in T-MEF MCMV group were up-regulated as the infection time extended,which were two times and three times of the control group level,respectively.The mRNA level(A value)of TGF-β in MCMV infected MEF increased from baseline of 1.09±0.13 to 3.15±0.54 on day 3 after infection.The expression of TGF-β in supernatant 3 days after infection was(3.85±0.32) μg/L,which was significantly higher than that before infection[(1.74±0.14)μg/L,P<0.05].Conclusions Activated T cells have antiviral effect.However,the function of T cells is rapidly inhibited after activation,which may be due to the expression of Foxp3 mRNA induced by MCMV infected MEF and increased CD4+ CD25+ Treg proportion of the co-cultured T cells.TGF-β level is significantly increased after CMV infection,which may be an important mechanism of Treg proliferation.MCMV may manipulate Treg to evade specific immune elimination and,as a result,to cause CMV replication.
4.Decreased expression of γ-aminobutyric acid receptor in the hippocampal tissues of pharmacoresistant temporal lobe epileptic rats
Guofeng WU ; Jing SHI ; Zhen HONG ; Feng ZHOU
Chinese Journal of Neurology 2013;46(10):702-705
Objective To establish a multi-drug resistant model of temporal lobe epilepsy,and to observe the changes of γ-aminobutyric acid (GABA) receptor expression in the hippocampal tissues so as to explore its effects in pharmacoresistant epileptogenesis.Methods One hundred rats were selected to prepare the amygdaloid kindled model of epilepsy by chronic stimulation of amygaloid basal lateral nucleus.After the kindled model of epilepsy was prepared successfully(n =52),pharmacoresistant epileptic rats were selected according to their response to the phenobabital and phenytoin.The selected pharmacoresistant epileptic rats (n =8)were sacrificed and the hippocampus was removed to determine the GABA receptor expression,and the same number of pharmacosensitive epileptic rats was used as control.Results The pharmacoresistant epileptic rats displayed degenerative and necrotic hippocampal neurons.The arrangement of hippocampal neurons was disordered,and the structural characteristics of the arrangement of the hippocampal neurons disappeared.The gray values of GABAA-positive neurons in the hippocampal tissues (141.15 ± 14.72) increased significantly compared with the pharmacosensitive epileptic rats (92.56 ± 5.17; t =3.380,P =0.006).Western blot method demonstrated that the band of GABAA became narrowed and thin.The relative quantity of GABAA in the hippocampal tissues (0.38 ± 0.08) decreased significantly as compared with the pharmacosensitive epileptic rats (0.88 ± 0.18).A significant difference was observed (t =5.420,P =0.002).Conclusions GABA receptor expression might be decreased in the hippocampal tissues of pharmacoresistant epileptic rats.It might play a certain role in the formation of pharnmacoresistant epilepsy.
5.Effect of hippocampal electrical stimulation on the sodium channel current in CA1 region neurons in rats with pharmacoresistant temporal lobe epilepsy
Guofeng WU ; Zhen HONG ; Binli GAO ; Feng ZHOU
Chinese Journal of Neurology 2013;46(8):513-518
Objective To establish a multi-drug resistant model of temporal lobe epilepsy,and then the sodium current of pyramidal neurons in CA1 areas of the hippocampus was used as as index to observe the effect of hippocampal stimulation on pharmacoresistant epileptic rats.Methods Eighty Wistar rats were selected to prepare an amygdaloid kindled model of epilepsy by chronic stimulation of amygaloid basal lateral nucleus.When the kindled model of epilepsy was prepared successfully,the pharmacoresistant epileptic rats were selected according their response to phenobabital and phenytoin.The selected pharmacoresistant epileptic rats were divided into a hippocampal stimulation group (HS group) and a pharmacoresistant control group (PR group).A low-frequency hippocampal stimulation was performed in the HS group,while the PR group received sham stimulation.The whole-cell recording technique by patch-clamp was used to observe the changes of sodium current of hippocampal pyramidal neurons after the hippocampal stimulation.Results Compared with the PR group,the pharmacoresistant epileptic rats in HS group underwent low-frequency stimulation for 2 weeks showed that the amygdale stimulus-induced seizures were decreased (2.32 ± 0.38 in HS group and 4.45 ± 0.42 in PR group,t =84.600,P =0.000) and the parameters of the after-discharges were improved significantly.In HS group,the peak current shifted towards depolarization,the sodium channels were difficult to activate,and were more susceptible to inactivation.Moreover,the recovery time after the sodium channel inactivation was slower in HS group ((17.9 ±0.6) s) than in PR group((16.3 +0.3) s,t =-25.420,P =0.000).Conclusions Hippocampal stimulation may inhibit the sodium channel current of pyramidal neurons in CA1 areas of hippocampus.The mechanism of hippocampal stimulation in the treatment of pharmacoresistant epilepsy might be achieved partly by inhibiting the sodium channel current so as to decrease the excitability of hippocampal neurons.
6.A system review of randomized controlled trials on treating chronic stable angina by rhodiola.
Jian-Feng CHU ; Guang-Wen WU ; Guo-Hua ZHENG ; Feng ZHENG ; Jian-Feng XU ; Jun PENG ; Zhen-Feng HONG
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(8):940-946
OBJECTIVETo systematically assess the efficacy and safety of Rhodiola in treating chronic stable angina pectoris.
METHODSOur group searched the Cochrane library, PubMed, Embase, Chinese biomedical literature database (CBM), VIP database (VIP), Chinese Journal Full-text Database (CNKI) for the literature published in English and Chinese till April 2013. Randomized controlled trials (RCTs) were included on the therapeutic effect of Rhodiola or Rhodiola plus conventional Western medicine in comparison with the conventional Western medicine treatment on stable angina. Data were extracted according the data extraction form. The literature methodological quality was assessed by using the Cochrane handbook, and data analyzed by Rev-Man 5.2 Software for Meta-analysis. The effect indicators of outcomes was expressed by odds ratio (OR) and 95% CI.
RESULTSA total of 7 randomized controlled trials, 662 cases of stable angina pectoris patients met the inclusion criteria and all published in Chinese, without one scientific design and high quality literature. Compared with the conventional Western medicine treatment, combined with oral administration of Rhodiola could improve the efficiency of anti-angina (OR = 2.49, 95% CI: 1.02 - 6.09). Combined with intravenous infusion of Rhodiola could also improve the efficacy of angina pectoris (OR = 4.86, 95% CI: 2.4 - 9.82). Oral administration of Rhodiola couldn't improve ECG efficacy (OR = 1.25, 95% CI: 0.67 - 2.34). Intravenous infusion of Rhodiola could improve the clinical efficacy (OR = 2.94, 95% CI: 1.61 - 5.35). Combined with the conventional treatment, intravenous infusion of Rhodiola could improve the whole blood viscosity (low and high shear rates) and inverse variance (IV) (-1.36 and -0.99, 95% CI: -1.65 - 1.07 and -1.26 - 0.71), but could not reduce serum fibrinogen and D-dimer level. The incidence rate of adverse reactions was higher than that of the conventional treatment combined with Rhodiola (OR = 0.1, 95% CI: 0.02 - 0.51).
CONCLUSIONSOn the basis of routine treatment, Rhodiola could further improve patients' symptoms. Combined with intravenous medication, Rhodiola could increase the ECG improvement rate, and reduce adverse reactions. But the methodological quality of included studies was poor, the number of samples was small, and influence factors such as the intervention period was short. This conclusion needs scientific and rational design in a larger sample, multicenter clinical trial to verify.
Angina, Stable ; drug therapy ; Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Randomized Controlled Trials as Topic ; Rhodiola ; Treatment Outcome
7.Advancements in stabilization technologies for membrane protein and its application in drug screening
Jia-hao FANG ; Yu-hong CAO ; Yu-zhen HE ; Zhan-ying HONG ; Yi-feng CHAI
Acta Pharmaceutica Sinica 2021;56(9):2325-2334
Membrane proteins are the main undertakers of biofilm function, and also the most important target group for innovative drug discovery and research. About 60% of drugs targets are membrane proteins. Due to the obvious aggregation and denaturation tendency of membrane proteins in aqueous solution, it is difficult to simulate the membrane like environment to maintain the correct conformation of membrane proteins
8.Effect of hemodialysis with plasma-based dialysate plus high volume hemofiltration on plasma cytokines in patients with liver failure
Wei CHEN ; Hong-Bao LIU ; Zhen-Jiang LI ; Ke-Feng DOU ; Zhen-Shun SONG ; Yue-Qing XU ; Han-Min WANG ;
Chinese Journal of Nephrology 2005;0(12):-
Objective To propose a new blood purification modality-hemodialysis with plasma- based dialysate (HD-PBD) plus high volume hemofiltration (HVHF) for patients with liver failure, and to evaluate the effect of this treatment on plasma cytokines.Methods Twelve patients with liver failure were included in this study.All patients received HD-PBD therapy in the first 6 hours,and then were treated with HVHF for 24 hours with the same filter (AV600).The levels of TNF-?,IL-1?, IL-6 and IL-8 in plasma before and after HD-PBD plus HVHF for 6 and 24 hours were examined respectively by ELISA,and changes of clinical parameters were observed at the same time point. Serum bilirubin,total bile acids (TBA),serum ammonia,blood urea nitrogen (BUN) and serum creatinine (Scr) were detected before and after treatment.Arterial blood gas analysis and the concentration of electrolytes were monitored before and after treatment.Results (1)HD-PBD for 6 hours was more effective than HVHF for 24 hours in removal of serum bilirubin and TBA(P<0.05). (2)Serum ammonia,BUN,Ser,arterial blood HCO_3~-,PCO_2,PO_2 and electrolytes did not show significant difference before and after HD-PBD (P>0.05),but these parameters significantly changed before and after HVHF (P<0.05).(3)The average level of serum bilirubin was sharply decreased after HVHF for 24 h following HD-PBD(P<0.05).(4)After HD-PBD plus HVHF,there was a marked reduction of the plasma levels of TNF-?,IL-6 and IL-8.Conclusions HD-PBD plus HVHF,a newly proposed modality for patients with liver failure,can effectively decrease serum bilirubin,TBA,BUN,Scr,ammonia and cytokines,and adjust water-electrolyte as well as acid- alkali balance.It is a low-cost,safe,simple and convenient therapy.
9.Effect of Kangquan Recipe on apoptosis regulatory genes bax and bcl-2 mRNA in prostate of rats.
Yuan-peng HUANG ; Jian DU ; Zhen-feng HONG
Chinese Journal of Integrated Traditional and Western Medicine 2007;27(8):711-714
OBJECTIVETo investigate the effect of Kangquan Recipe (KQR) on apoptosis regulatory genes bax and bcl-2 mRNA in prostate of rats.
METHODSBenign prostatic hyperplasia model rat was established by injecting testosterone after castration. The model rats were killed and prostate glands were removed for examination after being treated with administration of KQR by gastrogavage for 30 days. The wet weight of prostate was measured and the mRNA expressions of bax and bcl-2 in rats' tissue of abdominal lobe of prostate were determined by RT-PCR.
RESULTSCompared with the model group, wet weight of prostate was lower significantly in the groups treated with different dosages of KQR (P < 0.05 or P < 0.01), and that in the high dose KQR treated group was similar to that in the normal group (P > 0.05). Compared with the model group, the expressions of bax mRNA and ratios of bax/bcl-2 were significantly higher and the expressions of bcl-2 mRNA significantly lower in the KQR treated groups (P < 0.01), and these indexes in the high dose KQR treated group were insignificantly different from those in the normal group (P > 0.05).
CONCLUSIONKQR shows an obvious treatment effect on rats with benign prostatic hyperplasia, the mechanism might be through effectively regulating the expressions of bax mRNA and bcl-2 mRNA in prostatic tissue to accelerate the cell apoptosis of prostate in obvious dose-effect manner.
Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; therapeutic use ; Gene Expression ; drug effects ; Male ; Phytotherapy ; Prostate ; drug effects ; metabolism ; pathology ; Prostatic Hyperplasia ; chemically induced ; drug therapy ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Testosterone ; bcl-2-Associated X Protein ; genetics
10.Case-control study on effects of vacuum drainage on perioperative blood loss after total hip arthroplasty for the treatment of femoral neck fractures.
Guo-Gang LUO ; Hong-Zhen ZHANG ; Jian-Chuan YAO ; Zhong-Qin LIN ; Hai-Feng XIE
China Journal of Orthopaedics and Traumatology 2015;28(3):210-213
OBJECTIVETo compare postoperative blood loss under different negative pressures of drainage after total hip arthroplasty for the treatment of femoral neck fractures.
METHODSFrom January 1st to December 30th 2013, 74 patients with femoral neck fractures treated with total hip arthroplasty were randomly divided into two groups: high negative pressure drainage group and low negative pressure drainage group. In high negative pressure drainage group, there were 34 cases including 10 males and 24 females, with a mean age of (75.94 ± 9.02) years old, and the patients were treated with 60 kPa negative pressure of drainage. In the low negative pressure drainage group, there were 40 cases including 13 males and 27 females, with an average age of (74.93 ± 8.90) years old, and the patients were treated with 30 kPa negative pressure of drainage. The amount of total drainage, total blood loss, and hemoglobin change were compared between these two groups.
RESULTSAll the patients got primary healing without infections. In high negative pressure drainage group,the change of hemoglobin was (41.74 ± 15.69) g/L, total blood loss was (1,217.73 ± 459.50) ml and the drainage volume was (312.94 ± 103.44) ml; while in low negative pressure drainage group,the results were (34.90 ± 12.90) g/L, (904.01 ± 381.58) ml and (129.25 ± 44.25) ml separately. All the results in high negative pressure drainage group were higher than those in the other group. Three days after operation, the change of hemoglobin was (46.00 ± 13.29) g/L and total blood loss was (1,304.72 ± 421.75) ml; while in low negative pressure drainage group, the changes of hemoglobin was (43.87 ± 11.39) g/L and total blood loss was (1,196.78 ± 344.20) ml; there were no statistically significant differences between two groups.
CONCLUSIONWhen placing drainage devices after total hip arthroplasty for the treatment of femoral neck fractures, the level of negative pressure should be chosen according to preoperative level of hemoglobin and HCT in patients. For old patients with femoral neck fracture, low negative pressure is more suitable.
Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; methods ; Case-Control Studies ; Female ; Femoral Neck Fractures ; surgery ; Humans ; Male ; Middle Aged ; Negative-Pressure Wound Therapy ; Postoperative Hemorrhage ; prevention & control