Objective To evaluate the inhibitory effect of the small interfering RNA(siRNA) on hepatitis B virus(HBV)in vivo which targets HBV S gene region.Methods An animal model of HBV infection was developed hydrodynamically by injecting pcDNA3.1-HBV together with siRNA through the tail vein of Balb/c.HBsAg was analyzed by time resolved immunofluorometric assay, HBV DNA was analyzed by fluorogenic quantitative PCR(FQ-PCR),HBV S-mRNA was detected by semi-quantitative RT-PCR,and viral specific proteins(HBsAg and HBcAg)in the liver were assayed by immunohistochemical staining.Results In the mice,the siRNA could effectively inhibit the secre- tion of HBsAg,reduce the titers of HBV DNA,and immunohistochemical results also indicated that the number of HBsAg and HBcAg positive cells was reduced.The inhibitory effect of siRNA on HBV lasted 3 clays at least.Conclusion These results demonstrate that the siRNA targeting HBV S gene region can substantially and specifically inhibit HBV replication and expression in vivo.

Objective To investigate effect of mild hypothermia on changes of somatosensory evoked potential (SEP) and synaptophysin mRNA level after traumatic brain injury (TBI) and determine hypothermia-induced neuroprotection.Methods Forty-five SD rats were allocated into mild hypothermia group,TBI group and sham operation group with 15 rats per group according to the random number table.Left-side fluid percussion impact was performed to induce models of TBI.Rats were exposed to hypothermia environment (32-35℃) for 6 hours in mild hypothermia group after TBI.Rats in sham operation group were treated by only drilling on left side of the head,rather than hitting.To evaluate function outcome,modified neurological severity score (mNSS),SEP and synaptophysin mRNA level were measured at 6 hours,24 hours and 7 days postinjury.Results The mNSS in mild hypothermia group lowered compared with TBI group,especially at 24 hours and 7 days (P ＜ 0.05).SEP in mild hypothermia group was significantly shortened at 6 and 24 hours compared with TBI group (P ＜ 0.05),but SEP revealed no significant difference among the 3 groups at 7 days (P ＞ 0.05).Level of synaptophysin mRNA in mild hypothermia group increased at 6 hours postinjury compared with TBI group [(0.08 ± 0.02) vs (0.12 ±0.04)],with further increase at 7 days postinjury[(0.06 ± 0.01) vs (0.33 ± 0.10)] (P ＜0.05).Conclusion The shortage of nerve conduction time of the injured side and promotion of nerve regeneration suggest the neuroprotective role of mild hypothermia following TBI.

Objective To investigate the effect of sodium fluoride(NaF) on proliferation, differentiation and the mRNA expression of osteoprotegerin (OPG) and receptor activator of nuclear factor κβ ligand (RAN KL) of mouse osteoblasts. Methods Osteoblasts were isolated from calvarias of Kunming mice born in 1 - 2 d and cultured. Various concentrations of NaF(0, 10-8, 10-7, 10-6, 10-5, 10-4, 10-3mol/L) were added to the culture medium, the proliferation and activity of alkaline phosphatase(ALP) was determined after 72 h or 120 h. The expression of OPG mRNA and RANKL mRNA was analyzed by semi-quantification RT-PCR. Difference among groups was analyzed by One-Way AN0VA. Difference between two groups was analyzed by LSD-t test. Results There was significant difference in cell proliferation among groups after 72 h(F = 13.806, P < 0.05). Compared with control group(0.434 ± 0.010) , the proliferation was significantly induced in 10-7 - 10-4 mol/L groups treated osteoblasts (0.448 ± 0.010, 0.453 ± 0.013, 0.454 ± 0.016, 0.449 ± 0.018, all P< 0.05), and was significantly suppressed in 10-3 mol/L group(0.401 ± 0.009, P < 0.05). There was statistic difference in the activity of ALP among groups(F = 9.021, P < 0.05). Compared with control group (1.677 ± 0.682), the activity of ALP significantly increased in 10-7 - 10-5 mol/L groups[ (2.447 ± 0.756) × 106, (2603 ± 0.183) × 106, (2.687 ± 0.886) × 106 U/L, P < 0.05 or P < 0.01 ] and significantly decreased in 10-4 mol/L group[ (1.479 ± 0.366) × 106 U/L, P < 0.05 ]. There was significant difference in the expression of OPG mRNA among groups(F = 11.299, P< 0.05). Compared with control group (1.000 ± 0.000), the expression of OPG mRNA was significantly increased in 10-7 - 10-4 mol/L groups( 1.058 ± 0.027, 1.053 ± 0.026, 1.088 ± 0.055, 1.069 ± 0.008, P < 0.05 or P < 0.01) , while significantly decreased in 10-3 mol/L group (0.941 ± 0.029, P< 0.05). There was no difference in RANKL mRNA expression among groups (F= 1.311, P> 0.05). The ratio of RANKL/OPG decreased with increasing doses of fluoride and increased in 10-4, 10-3 mol/L groups, but there was no difference between groups(F = 1.376, P> 0.05). Conclusions A biphasic pattern of proliferation and differentiation has been induced in mouse osteoblasts, which manifests stimulation effect in low doses and suppression in higher doses. Low doses of sodium fluoride suppress differentiation and maturation of osteoblasts by increasing expression of OPG mRNA, while high doses of sodium fluoride enhance differentiation and maturation of osteoblasts by decreasing expression of OPG mRNA.

Several kinds of column chromatography method were used to investigate the chemical constituents of the leaves of Lophatherum gracile. The structures of the isolated compounds were identified based on their physicochemical properties and spectral data. A new flavone C-glycoside was isolated and its structure was identified as 3'-methoxyl-luteolin 6-C-beta-D-galactopyranosiduronic acid (1 --> 2) -alpha-L-arabinopyranoside (1).
Antiviral Agents ; chemistry ; isolation & purification ; Flavones ; chemistry ; Glycosides ; chemistry ; isolation & purification ; Hydrolysis ; Plant Leaves ; chemistry ; Poaceae ; chemistry

Objective To investigate the effect of taurine transporter in the process of protection of brain edema in rats with severe traumatic head injury. Methods A total of 24 Male Sprague-Dawley rats were randomly divided into 4 groups. Except the control rats (Group Sham), all other three groups were subjected to lateral fluid percussion head injury. The TBI (Traumatic brain injury) models (Group TBI) and surgical control rats (Group Sham) were injected with saline through caudal vein after surgery, while the Taurine prevention and Taurine treatment models (Group Pre Tau and Group Tau) were injected with 120 g/L taurine solution before or after surgeries respectively. Water content in each brain, mRNA and protein expres?sion of aquaporin 4 and taurine transporter in the injured rat brain hemispheres were all evaluated over the time course of the study (7 d) in each group. Results Compared with rats in Group Sham, water content in each brain increase, mRNA tran?scription and protein expression of AQP4 were both up regulated but the mRNA transcription and protein expression of TauT were both down-regulated in rats in TBI group. Compared with rats in TBI group, brain water content, mRNA transcription and protein expression of AQP4 all decrease while mRNA transcription and protein expression of TauT all increase in rats in Pre tau and Tau groups. There is no statistical difference of TauT expression between rats in pre-tau group and Tau group. Conclusion Taurine exert its neuron protection role through draining water content from brain and down regulating expres?sion of AQP4 but rising expression of TauT after TBI.

Objective To investigate effect of taurine on respiration chain enzyme activity of mitochondria 24 hours after severe traumatic brain injury (TBI) in rats.Methods Fifty-six SD rats were divided into sham group,TBI group,taurine treatment group,and taurine prevention group according to the random number table,with 14 rats per group.Fluid percussion brain injury models were used.Via the caudal vein,normal saline was administered to animals in sham and traumatic brain injury groups immediately after injury,while taurine (200 mg/kg)was administered to animals in taurine treatment group after injury and in taurine prevention group 4 days before injury.Brains were harvested 24 hours postinjury for assays of HE staining and electron microscopy.Mitochondrial respiratory chain complex Ⅰ-Ⅴ activities were detected.Results TBI group presented swelling neurocytes,cell loss,karyopyknosis,shortened even vanished process,and inflammation cell infiltration at the edge of necrosis in HE staining.By contrast,morphological improvement was significant in taurine treatment group but only some neurons were intact in taurine prevention group.Swelling mitochondria and broken or vanished mitochondrial crests were seen in TBI group under the electron microscope.However,normal or minor swelling mitochondria was seen in taurine treatment group and cytoplasm slightly porous and absence of mitochondrial crests were seen in taurine prevention group.Activities of complex Ⅰ,Ⅱ and Ⅴ were significant lower in TBI group (32.52±2.41,4.68 ±0.15,2.49 ±0.73) compared to those in sham group (34.03 ±0.46,5.04 ±0.29,3.20±0.68) and in taurine treatment group (33.95±0.85,5.12-±0.23,3.53 ±0.48) (P＜0.05).And complex Ⅰ in taurine prevention group was significantly enhanced as well (34.44 ± 0.36,P ＜ 0.05).Conclusion Taurine may protect the brain tissues and mitochondrial structure from impairment in TBI rats by improving mitochondrial enzymes activity and reducing secondary energy loss.

Objective To investigate the expression level of adenosine 2A receptors in peripheral blood lymphocytes and the correlation with disease progression of Parkinson's disease (PD).Methods In this retrospcctive study,out patients with PD(42 cases)and healthy controls(32 cases) were recruited at Beijing Hospital.The expression level of A2A receptors in peripheral blood lymphocytes was detected by flow cytometry.The concentration of free A2A receptors in plasma was detected by enzyme linked immunosorbent assay (ELISA).The expression of A2A receptors and plasma free A2A receptors in peripheral blood lymphocytes of the PD group and the control group was compared.Multivariate regression analysis and single factor correlation analysis were performed on sex,age,course of disease,duration of medication,drug type and dose,motor complications,and PD unified Parkinson's disease rating scale (UPDRS)score.Results A2A receptor expression in lymphocytes was significantly higher in the PD group than in the control group (8.96 ± 4.73)％ vs.(5.39±2.42)％ (t=4.210,P＜0.05).There was no significant difference in A2A receptor concentrations in plasma between the two groups (1.82 ± 1.91) μg/L vs.(1.15 ± 0.71) μg/L(t=1.078,P＞0.05).In the PD group,A2A receptor expression in lymphocytes in patients with motor complications was statistically lower than in patients without them (P＜ 0.05).Lymphocyte A2A receptor levels in the 5-9 years duration subgroup were significantly lower than those in the ＜5 years duration subgroup (Z=2.780,P＜0.01) and the≥10 years duration subgroup (Z=-2.149,P＜0.05).Conclusions The expression of A2A receptors in peripheral blood lymphocytes is correlated with PD.The expression of A2 A receptors in peripheral blood lymphocytes of patients with PD fluctuates with the occurrence of motor complications and the progression of disease.Further research is needed to establish A2A receptors as a biomarker for monitoring disease progression.

ObjectiveTo detect the levels of bone morphogenetic protein-2(BMP-2) mRNA in gunshot fracture healing of the rabbits.MethodsA model of primary treating gunshot fracture of the rabbit with external fixator and the real time fluorescence quantitative reverse transcription polymerase chain reaction(FQ-RT-PCR) technology with SYBR Green I were established.The levels of BMP-2 mRNA 1,2,3 and 4 weeks after operation were detected with FQ-RT-PCR respectively.ResultsIn the control group,BMP-2 mRNA of the bone tissue began to rise at the first week after fracture and the peak of mRNA expression appeared at the second week.The expression returned to normal at the forth week.In the experimental group,the BMP-2 mRNA rose more slowly that it arrived peak stage at the third week and was significantly lower in experimental group than that in control group at the same phases.ConclusionReal time FQ-RT-PCR is a good method for measuring the levels of BMP-2 mRNA during gunshot fracture healing.The mRNA expression rose more slowly and was significantly lower in gunshot fracture than in common fracture at the same phases.

Background and purpose: The incidence of breast cancer increases as patients age, elderly patients account for a large proportion. Due to the insufifcient systemic therapy, more complications and poorly physical condition, the prognosis of elderly patients is often worse than the younger. The aim of this study was to investigate the safety and tolerance with non-anthracyclin regimen as adjuvant chemotherapy in elderly breast cancer patients. Methods:From Nov. 2008 to Jan. 2012, 56 patients (≥65 years) after surgical excision were enrolled into this study. The patients were divided into two groups:TC and PC groups. Each patient received 4 or 6 cycles of chemotherapy of PC (175 and 600 mg/m2, respectively;n=21) or TC (75 and 600 mg/m2, respectively;n=35), administered intravenously every 3 weeks, as adjuvant chemotherapy. Radiation therapy (as indicated) and endocrine therapy, for patients with hormone receptor-positive disease, were administered after completion of chemotherapy. Results: In this study, 50 patients completed chemotherapy as plan, the proportion of two groups were above 90%. After a median follow-up of 33 months, the median disease-free survival(DFS) and overall survival(OS) were not reached. The relapse-free rate and survival rate were 89.5%and 100%in the PC regime group, which were 90.3%and 96.8%in the TC regime group. Major toxicities included:neutropenia, leucopenia, alopecia, nausea, vomiting and various degree of peripheral neuropathy. The incidence of gradeⅢ-Ⅳneutropenia was 76.2%in PC group vs 48.6%in TC group (P=0.044). The most common cause for withdrawing from treatment was to be unable to tolerate the adverse effects. Conclusion:Adjuvant chemotherapy with paclitaxel and cyclophosphamide is safe, tolerable and promising for elderly breast cancer patients.

Objective To study the effects of dynamic,support-inducing exercise on the support,balance and gait ability of patients with moderate-to-severe hemiplegia after stroke.Methods Fourteen stroke patients were randomly assigned to an experimental or a control group (7 cases to each).The patients in the experimental group received both dynamic,support-inducing exercise and routine exercises,while the patients in the control group received routine exercises only.Before training and after 40 and 60 days of training,their functional capacity was evaluated with the Chinese stroke scale (CSS) for neurological deficits,Berg's balance scale (BBS) and using functional ambulation categories (FACs).Results Before training there was no inter-group difference in average CSS or BBS scores or in FACs.For the experimental group there were significant intra-group differences compared with 0th day in all three items at both time points.At days 40 and 60 there were also significant intra-group differences in BBS scores and FACs in the control group,and CSS scores improved significantly only in the experimental group.At day 40 there were significant inter-group differences in average CSS,BBS and FAC results.However,by day 60 a significant difference persisted only in average CSS scores.Conclusions Dynamic,support-inducing exercise can improve support,balance and gait in patients with moderate-to-severe hemiplegia after stroke.