Objective To investigate the effect of cyclosporine A(CsA)on viral protein syn- thesis and hepatitis B virus(HBV)DNA replication in vitro.Methods The HBV DNA transfected cell line HepG2.2.15 was treated with different concentration of CsA(0.6-20.0?g/ml)for 4 days. Hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)in supernatant were detected by enzyme-linked immunosorbentassay(ELISA);intracellular hepatitis B core antigen(HBcAg)mR- NA and HBV DNA were analyzed by RT-PCR and slot blot hybridization,respectively;the phospho- rylation at tyrosine acid position 402 of PyK2 kinase(PyK2 Y402)was detected by Western blot.Re- sults CsA could suppress the expression of HBsAg and HBeAg,and inhibit the HBV DNA replica- tion in a dose-dependent manner.The suppression rate of HBsAg and HBeAg under the action of CsA at the concentration of 10.0?g/ml for 4 days was 49.7% and 34.3%,respective;similar effect was observed on HBV DNA replication,HBV DNA was only 34.9% of the control at the concentration of 10.0?g/ml of CsA.The phosphorylation level of PyK2 Y402 was declined under the action of CsA at the concentration of 2.0?g/ml.Conclusions CsA can inhibit the expression of HBsAg,HBeAg and HBV DNA replication in the HepG2.2.15 cell line in a dose-dependent manner.Suppression of the phosphorylation level of PyK2 Y402 maybe involved in the mechanism of the inhibitory activity of CsA on HBV replication.

AIM: To investigate the effect of the heat shock response on the reperfusion arrhythmias(RAs) and the possible mechanism involved. METHODS: Fifty-five Sprague Dawley rats were randomly divided into 2 groups: the heat shock group (group H, n=29 ) and the control group (group C, n=26 ). The rats in group H were preconditioned with heat shock 24 hours before, and that in group C were not. The hearts of 16 rats in group H and 16 in group C were exercised and mounted on a non-circulating Langendorff perfusion apparatus and perfused retrogradely with modified K-H buffer and mimic ischemia/reperfusion was applied. Additionally, conventional intracellular microelectrode techniques were used for recording such electrophysiological parameters as resting potential(RP), action potential amplitude(APA), over shot(OS), maximum depolarization velocity(Vmax) of the hearts of other 13 rats in group H and 10 in group C. RESULTS: ①Prior heat stress significantly decreased reperfusion arrhythmia. ②The amount of CK release in the effluent in group H was much less than that in group C. ③Myocardial HSP70 content was elevated significantly in group H. ④Heat stress significantly increased myocardial anti-oxydases activity and decreased lipid peroxydative products. Additionally, heat stress significantly reduced the Vmax of action potential. It indicated that rapid Na + channel of papillary muscles may be inhibited by heat shock. The degree of change of Vmax after ischemia in H group was significantly less than that in group C. And the time of reperfusoin with Tyrode's solution till the action potential appeared as large as that before perfusion with mimic ischemic solution is shorter in group H than in group C. CONCLUSION: Heat shock pretreatment markedly reduces ischemia/reperfusion-induced injury of heart and ventricular arrhythmias in rats and this effect may be associated with the inhibition of rapid Na + channel of papillary muscles by heat shock and the increase in myocardial HSP70 and anti-oxydase activity.

BACKGROUND: Migu tablet, a Chinese drug for kidney invigorating, is effective on preventing and treating osteoporosis, but the concrete mechanism of pharmacology is still not clear. Transforming growth factor-β1(TGF-β1) is an important cytokine, which can regulate bone resorption and formation.OBJECTIVE: To investigate the effect of kidney invigorating recipe on mRNA expression of TGF-β1 of osteoblast.DESIGN: A completely randomized controlled study was conducted.SETTING: Department of Traumatic Orthopedics, Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: This experiment was conducted at the laboratory for bone metabolism of integration of Chinese and western medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from May 2003 to April 2004. Experimental rats: Totally 16 newborn SD rats of clean degree were involved. Experimental drug: Medical liquor of Migu tablet was prepared in the Department of Traumatic Orthopedics,Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology. The subscription was mainly composed of Chinese herbs such as Herba Epimedii, Cortex Eucommiae, Semen Juglandis,Radix Rehmanniae, Radix Achyranthis Bidentatae. and so on. Positive control drug: which was recombinant basic fibroblastic growth factor (rbFGF), was purchased from Beijing Banding Company. Negative control group was subdivided into negative control of probe and antibody METHODS: 100,1 000,5 000,10 000 mg/L Chinese herb Migu tablet liquor for kidney invigorating and positive control drug 5 μg/L rbFGF were added into the osteoblasts of cranial bones of newly born SD rats separated and cultured in vitro. 24 hours later, nuclear acid molecular in situ hybridization of osteoblasts were analyzed by self-made digoxin-labeled TGF-β1 cDNA probe . The mean absorbance of positive particles representing the mRNA expression of TGF-β1. A total of 40 osteoblasts were randomly chosen from each group under 200-fold amplification. The average absorbance of hybridized particles of the cells was measured with TJTY-300 automatic image analyzer.MAIN OUTCOME MEASURES: mRNA expression of TGF-β1 in osteoblasts of each group.RESULTS: Automatic image analyzer showed that the TGF-β1 mRNA expressions of Migu liquor groups whose concentration were 5 000 mg/L and 10 000 mg/L were respectively 1.18 times and 1.30 times that of control group, with a significant difference. [The mean absorbance's of hybridized particles of the cells in the 5 000,10 000 mg/L Migu liquor groups and negative control were 0.213 67±0.015 00,0.237 03±0.021 73,0.181 27±0.015 28 ,respectively, P ＜ 0.05 and P ＜ 0.01].Although the mean absorbance ( 0.254 45±0.020 81)of the hybridized particles of the cells in the 5 μg/L recombinant rbFGF was higher than those of 5 000,10 000 mg/L Migu liquor groups, but there was no significant difference(P ＞ 0.05).CONCLUSION: Migu tablet for kidney invigorating can stimulate the secretion and synthesis of TGF-β1 in osteoblasts, thus promote bone formation and inhibit bone resorption.

BACKGROUND:The therapeutic effects of donkey-hide glue reinforcing bone oral solution on osteoporosis have been determined, but the exact effective mechanism is to be approached. OBJECTIVE: To investigate the effects of donkey-hide glue reinforcing bone oral solution (DGRBOS) medicated serum on osteoprotegerin (OPG)and its ligand(OPGL)mRNAexpression of osteoblast in fetal rats and explore the molecular mechanism of treating osteoporosis with DGRBOS. DESIGN: A randomized controlled trial. MATERIALS: The experiment was carried out from June 2003 to October 2004 in Bone Metabolic Laboratory of Department of Integrative Chinese and Western Medicine, Affiliated Hospital of Tongji Medical College,Huazhong University of Technology and Science. Totally 30 3-month-oldWistar rats (15 males and 15 females) were randomly divided into 3 groups, I.e. DGRBOS group, estrogen group and control group, with 10 rats in every group. 12 clean newborn SD rats were selected to isolate and cul ture osteoblast. METHODS: ①After intragastric administration for 7 days, medicated serum was prepared respectively from the three groups. ②Skull osteoblast isolated from newborn SD rats was made into single cell suspension, then after digestion and passage, the subcultured osteoblast cell was made into cell suspension. The cultured osteoblasts were divided into 5 groups and given equal volumes of drug liquor. The DGRBOS group was given DGRBOS-medicated serum at the concentration of 100, 500 and 1 000 g/L which was diluted by nutrient solution; the estrogen group was given tibolone-medicated serum of 100 and 1 000 g/L; the control group was givenonly culture fluid. Meanwhile every group was given calf serum (100 g/L) for further culture. ③The osteoblast proliferation was measured by antigenic MTT colorimetric analysis and 3H-TdR penetration method. The in tra-cellular BGP contents were evaluated by radioimmunity .The mRNA expression of OPG and RANKL in osteoblast was analyzed by Rt-PCR. ④ One-way analysis of variance was applied to compare data among groups. MAIN OUTCOME MEASURES: mRNA expression of OPG and PAN KL in osteoblasts from fetal rats after intervention by medicated serum ofDGRBOS or Livial. RESULTS: ①The osteoblast proliferation measured by antigenic MTT colorimetric analysis and 3H-TdR penetration method showed that the proliferation in the DGRBOS group and tibolone group was enhanced moresignificantly than that in the control group (P ＜ 0.05-0.01), and reached maximal effect at the concentration of 500 g/L (P ＜ 0.01), but when the concentration was over 500 g/L, the effect tended to saturate. The medicated serum with all concentrations from DGRBOS and estrogen groups could increase the contents of BGP in osteoblasts (P ＜ 0.05). ②The mRNA expression of OPG reached the peak when the DGRBOS medicated serum was 1 000 g/L, and was obviously higher than that at the concentration of 100 and 500 g/L (P ＜ 0.05). The expression in DGRBOS group at the concentration of 1 000 g/L and in the estrogen group at the concentration of 100 and 1 000 g/L was apparently higher than that of the control group (P ＜ 0.01). ③The mRNAexpression of RANKL was the highest in DGR BOS group with 1 000 g/L concentration, and was markedly lower than that of the concentration of 100 and 500 g/L (P ＜ 0.05). The expression in DGRBOS group at the concentration of 1 000 g/L and in the estrogen group at the concentration of 100 and 1 000 g/L was noticeably lower than that in the control group (P ＜ 0.01).CONCLUSION: ①The DGRBOS could remarkably enhance osteoblast proliferation in dose-dependent and a dose-saturable manner, and the effect was close to that of tibolone. ②Partial mechanism of DGRBOS in treating osteoporosis might be promoting osteoblast proliferation and regulating OPG/RANKL expression.

Objective To investigate the relationship between apolipoprotein E(Apo E) gene poly-morphism and mild cognitive impairment (MCI) in patients with type 2 diabetes mellitus (T2DM), and e-valuate the correlative risk factors. Method 40 cases of type 2 diabetes with MCI and 80 cases of type 2 diabetes without MCI were enrolled in this study. The polymorphism of the Apo E gene was detected by PCR-restriction fragment length polymorphism(PCR-RFLP). According to the clinical data such as course of disease, plasma glucose, plasma fat and body mass index (BMI), the independent risk factors of T2DM and MCI were analyzed by non-conditional logistic regression. Results The frequency of Apo E ε_4 allele in the group of type 2 diabetes with MCI was higher than that without MCI ( 25.0% vs 10. 0% ), and the difference had statistical significance( P < 0. 01 ). The indexes of the statistical significant difference be-twcen the two groups were age, course of disease, postprandial blood glucose ( P2BG), HBA1C, BMI,family history of T2DM, hypertension, diabetic retinopathy, diabetic peripheral neuropathy, Apo E gene. The independent risk factors included diabetic retinopathy ( OR = 3. 452, P < 0. 05 ), diabetic peripheral neuropathy( OR = 3. 252, P <0. 05), Ape E gene( OR = 2. 441, P < 0.01 ), HBA1C ( OR = 1. 372, P <0.05), P2BG(OR = 1. 194, P <0.05), age(OR = 1. 194, P <0.01) and course of disease(OR =1. 142, P <0. 05). Conclusion Apo E ε_4 allele has significant relationship with T2DM and MCI. The age, course of disease, control of plasma glucose, and microvascular complication of diabetes have relation-ship with the cognitive function.

Objective To investigate the relationship between alteration of gene in cyclic adenosine monophosphate(cAMP) signal transduction system in rats after myocardial damage and changes of cardiac function and ventricular remodeling. Methods Twenty eight male Wistar rats weighing 220 g to 250 g were randomly divided into three groups: acute myocardial damage group(AMD, n = 10), chronic myocardial damage group (CMD,n = 9 ) and sham-operation group (control, n = 9). Animal model of acute myocardial damage was established by ligation of rats left coronary artery in the AMD and the CMD groups. Rats in control group were treated similarly, except that the coronary suture was not tied. Hemodynamics and echocardiography were measured before rats were sacrificed 24 hours after operation in control and AMD groups but those in CMD groups were sacrificed 8 weeks later. Cadiocyte apoptosis were estimated by TUNEL method, cAMP levels in heart were tested by radioimmunity and the mRNA expressions for inducible cAMP early repressor (ICER), cAMP response element binding protein (CREB), phosphodiesterase 3A (PDE3A) and bcl-2 were assayed by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Results The difference of left ventricular end diastolic diameter (LVEDD), left ventricular end diastolic pressure(LVEDP), maximal rising and falling rate of ventricular pressure,left ventricular systolic pressure (LVSP), eject fraction (EF) and fraction shortening (FS) were statistically significant among the three groups(F = 285.9, 196.8, 83.2, 80.4, 54.9, 196.6, 95.2, all P ＜ 0.01). LVEDD[(7.03 ±0.28), (8.20 ± 0.27)mm] and LVEDP[(11.19 ± 2.89), (19.76 ± 3.34)mmHg] in AMD and CMD groups were significantly increased, compared with those in control group[ (5.05 ± 0.30)mm, (- 5.62 ± 3.01 )mmHg, all P ＜0.01 ]. While maximal rising rate[ (2964 ± 449), (2214 ± 434)mmHg/s] and falling rate[(- 2617 ± 441),(- 1891± 424)mmHg/s] of left ventricular pressure, LVSP[ (94.19 ± 4.03), (85.85 ± 6.39)mmHg], EF[ (41.6 ±5.9)%, (35.9 ± 4.1 )%] and FS[ (36.9 ± 4.6)%, (23.1 ± 4.9)%] of left ventricular in the two groups were lower than those in control[(4759 ± 406)mmHg/s, (- 4327 ± 388)mmHg/s, (116.29 ± 8.25)mmHg, (80.9 ± 5.6)%,(53.1 ± 4.3)%, all P ＜ 0.01 ]. These changes in CMD group were more significant than those in AMD groups(P ＜0.05 or P ＜ 0.01 ). The difference of apoptotic index, cAMP and expression of ICER, CREB, PDE3A mRNA and bcl-2 mRNA were statistically significant among the three groups(F= 172.5, 141.0, 540.8, 246.8, 165.1, 563.9,all P＜ 0.01 ). Apoptotic index[ (32.8 ± 4.2)‰, (18.4 ± 3.9)‰] and cAMP in heart[ (9.95 ± 0.30), (5.60 ± 0.25)nmol/kg] in AMD and CMD groups were increased compared to control group[ (3.9 ± 1.7)‰, (2.48 ± 0.29)nmol/kg,all P ＜ 0.01 ], and those in CMD group were lower than in AMD group(all P ＜ 0.01 ). Expression of ICER mRNA (1.434 ± 0.093, 0.942 ± 0.076) and CREB mRNA(5.70 ± 0.50, 2.64 ± 0.51) in AMD and CMD groups were higher, and expression of PDE3A mRNA(48.98 ± 8.14, 16.68 ± 8.46) were lower than those in control group (0.154 ± 0.063, 1.08 ± 0.35, 105.94 ± 12.61, all P ＜ 0.01 ). The three genes in CMD group were fewer than those in AMD group(all P ＜ 0.01 ). bcl-2 mRNA was up regulated in AMD group(4.55 ± 0.27) and was down regulated in CMD group(0.35 ± 0.15) compared to control(2.18 ± 0.30, all P＜ 0.01). Conclusions There is PDE3A-ICER positive-feedback loop leading to myocyte apoptosis and heart failure after myocardial damage. The downregulation of PDE3A mRNA observed in chronic myocardial damage may play a causative role in the progression of ventricular remodeling and heart failure, in part, by inducing ICER mRNA and promoting cardiac myocyte dysfunction.

Objective To evaluate the effectiveness of bioelectrical impedance analysis(BIA) interventions on volume status in hemodialysis patient. Methods Searched The Cochrane Library, PubMed, EMbase, EBsco, Wanfang Data, China Biology Medicine, China National Knowledge Infrastructure to collect clinical trials. The retrieval time was from database to July 15, 2016. The studies were selected according to the inclusion and exclusion criteria and were critically appraised by two reviewers. Then the data of included studies were extracted. Meta-analysis was conducted by using RevMan 5.3 software and descriptive analysis. Results Ten clinical trials were included six random control trial, one controlled clinical trail, three longitudinal study, involving 2009 patients. The results of Meta-analysis showed that the effectiveness of BIA interventions on volume management could improve blood pressure (P=0.001), protect the heart function(left ventricular mass index,P=0.002), compared with the clinical evaluation method, the BIA assessment of dry weight of hemodialysis patients could reduce hospitalization rate 14%. Descriptive analysis showed that BIA intervention volume management had certain advantages for the survival benefit of patients, but the current research was still few, had not yet come to a certain conclusion. Conclusions BIA can improve the management of the volume status and dry weight of hemodialysis patients, so as to it can improve the clinical benefit and survival benefit of patients.Better methods and guidelines for assessing DW and using BIA need to be developed.We propose that experienced HD nursing staff be trained in the use of the BIA to help monitor patient over hydration and approximate dry weight in consultation with the nephrologists responsible for the care of these patients so as to obviate excessive residual over hydration between nephrology reviews.

Objective To investigate the influence of thymosin beta 4 (Tβ4) with two different dosages on the expression of transforming growth factor beta (TGF-β) and connective tissue growth factor (CTGF) in rats with renal tubular interstitial fibrosis,and to further estimate the changes of renal tubular interstitial lesions.Methods Rat models of renal tubular interstitial fibrosis were established by unilateral ureteral occlusion (UUO).The male SD rats were randomly divided into 4 groups and 15 rats in each group:sham group,model group,treatment group with 1 mg/L Tβ4 and treatment group with 5 mg/L T34.Rats in sham group and model group were poured into the same amount of saline.The renal function and renal pathological changes were observed after the second week.The mRNA and protein expression of TGF-β and CTGF in renal tissues was tested by in-situ hybridization and Western blotting.Results Compared with that in sham group,the expression of TGF-β mRNA and its protein,CTGF mRNA and its protein was significantly higher in model group (all P ＜ 0.01).Compared with rats of model group,Tβ4 treatment rats had lower mRNA and protein expression of TGF-β and CTGF (all P ＜ 0.01),and the expression in treatment group with 5 mg/L Tβ4 was lower than that in treatment group with 1 mg/L Tβ4 (P ＜ 0.05).And the expression of TGF-β mRNA was positively correlated with CTGF mRNA expression (r=0.697,P ＜ 0.01).The 24 h total urinary protein and the area of renal tubular interstitial lesion in model group were significantly more than those in sham group,and also more than those in Tβ4 treatment group (all P ＜ 0.05).Tβ4 treatment attenuated kidney damage,and the effects in treatment group with 5 mg/L Tβ4 were better than those in treatment group with 1 mg/L Tβ4.No difference in serum creatinine and blood urea nitrogen was observed among 4 groups (all P ＞ 0.05).Conclusions Tβ4 treatment can inhibit the renal TGF-β and CTGF expression of rats with tubular interstitial fibrosis in a dose-dependent manner,and play a protective role in kidney.

Objective:To investigate the relationship between tumor necrosis factor-α (TNF-α)-308A/G gene polymorphism and mild cognitive impairment(MCI)in patients with type 2 diabetes mellitus(T2DM),and their correlative risk factors thereof.Methods:Forty cases of T2DM with MCI and 80 cases of T2DM without MCI were selected for this study.The polymorphism of the TNF-α-308A/G was detected by PCR-restriction fragment length polymorphism (PCR-RFLP).According to the clinical data,such as course of disease,plasma glucose,plasma fat and body mass index(BMI),the independent risk factors of T2DM and MCI were analyzed by non-conditional logistic regression.Results:The frequency of TNF-α2 allele was significantly higher in the group of T2DM with MCI than that without MCI (P＜0.01).The indexes of the statistical significant difference between the two groups were the age,course of disease,postprandial blood glucose(P2BG),glycosylated hemoglobin,body mass index,family history of T2DM,hypertension,diabetic retinopathy,diabetic peripheral neuropathy and TNF-α.The independent risk factors included TNF-α,diabetic peripheral neuropathy,diabetic retinopathy,age and P2BG.Conclusion:There is a significant relationship between TNF-α2 allele and T2DM with MCI.There is a significant relationship between the age,control of plasma glucose and microvaseular complication of T2DM with the cognitive funotion.

Staphylococcus aureus (S. aureus) is an important human pathogen which can cause a chronic condition with a high relapse rate despite the aggressive antimicrobial treatment. Recent studies showed that intracellular pattern recognition receptors (including NOD) in response to bacteria or bacterial products play a proinflammatory role by activating nuclear transcription factor-κB (NF-κB). But how NOD2 mediates the proinflammatory response to S. aureus in mast cells (MCs) is unclear. So, in this study, we attempted to examine the role of NOD2 in inflammatory responses of MCs to S. aureus. P815 cells (a mouse mast cell line) were cultured. Real-time PCR was used to detect the NOD2 mRNA expression in P815 cells during S. aureus infection. The siRNA against NOD2 gene was synthesized and transfected into S. aureus-infected P815 cells. By using the methods of ELISA and flow cytometry, the effects of NOD2 gene silencing on cell phagocytosis, cytokine secretion, NF-κB activation and cell apoptosis of the S. aureus-infected P815 cells were examined. It was found that S. aureus infection could increase the expression of NOD2 mRNA in P815 cells. NOD2 gene interference in P815 cells reduced the number of S. aureus engulfed by P815 cells, the level of cytokines and the activation of NF-κB. In addition, S. aureus could induce the apoptosis of P815 cells, but NOD2 gene silencing did not affect the cell apoptosis rate. Our data suggested that NOD2 plays a key role in pathogen recognition, signal transduction, and NF-κB activation in the inflammatory responses of MCs infected by S. aureus.