Objective To confirm that astrocytes from cerebral cortex of newborn rat can be the target cells of Hantaan virus (HTNV)and Seoul virus (SEOV)infection and to observe changes of astrocytes after different infection time. Methods Astrocytes were prepared from cerebral cortex of newborn rat, and then infected with HTNV and SEOV. The established virus infections were confirmed by detection of virus nucleocapsid protein (NP) and S segment RNA in astrocytes using double-label immunofluoreseence, Western-blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Results The astrocytes from cerebral cortex of newborn rat were cultured successfully in vitro, which could be infected by HTNV and SEOV. The number of infected astrocytes and the virus titer in the infected astrocytes kept on increasing along with the extended infection duration. Conclusions Astrocytes from cerebral cortex of newborn rat are the target cells for HTNV and SEOV infection. Then establishment of in vitro cultured astrocytes model for Hantaviruses infection will be helpful for the study on the pathogenesis of hemorrhagic fever with renal syndrome.

BACKGROUND:Osteoprotegerin and nitrogen monoxidum play a key role in the prevention and treatment of osteoporosis. However, the correlation of the two in inhibiting the proliferation and differentiation of osteoclasts remains unclear.
OBJECTIVE:To observe the effect of different doses of osteoprotegerin on nitric oxide production and endothelial nitric oxide synthase activity in osteoclasts.
METHODS:Tartrate resistant acid phophatase staining was used to test whether the induced cells are osteoclasts. Osteoclasts were divided into six groups:blank control group (no reagent);negative control group (DMEM);four osteoprotegerin groups (10, 25, 50, 75μg/L of osteoprotegerin). Annexinv-FITC kit and flow cytometry were used to test the apoptosis rate of osteoclasts. Real-time PCR was used to detect the expression of osteoclasts marker gene, TRAP mRNA and protein kinase K mRNA. Nitric oxide production and nitric oxide synthase activity were determined using the corresponding kits. Four osteoprotegerin groups were added with L-NAME, a kind of nitric oxide synthase inhibitor, to test the changes of the apoptosis rates of osteoclasts and the changes of the expression of TRAP mRNA and protein kinase K mRNA of osteoclasts.
RESULTS AND CONCLUSION:Osteoprotegerin inhibited the differentiation of osteoclasts and induced the apoptosis. Osteoprotegerin concentration had a positive correlation with the apoptosis rate of osteoclasts, and a negative correlation with the numbers of osteoclasts and expression of marker gene TRAP mRNA and protein kinase K mRNA in osteoclasts. Osteoprotegerin boosted the nitric oxide production and endothelial nitric oxide synthase activity, and osteoprotegerin concentration was positively correlated to the nitric oxide production and endothelial nitric oxide synthase activity. After Raw264.7 cells were in vitro cultured, osteoprotegerin and nitric oxide play a synergic role in inhibiting osteoclasts production and promoting the apoptosis. We speculate that there is osteoprotegerin/endothelial nitric oxide synthase/nitric oxide signal pathway.

Filamentous fungi have been used in industrial fermentation extensively. Based on non-phosphorylating electron transport process, alternative respiration pathway (ARP) acts as an energy overflow, which can balance carbon metabolism and electron transport, allow the continuance of tricarboxylic acid cycle without the formation of ATP, and permit the turnover of carbon skeletons. Alternative respiration pathway also plays an important role in the stress response of fungi and the physiological function of conditioned pathogen. Alternative oxidase (AOX) is the terminal oxidase responsible for the activity of alternative respiration pathway, which exists widely in higher plants, parts of fungi and algae. Owing to the property that alternative oxidase (AOX) is sensitive to salicylhydroxamic acid (SHAM) and insensitive to conventional inhibitors of cytochrome respiration, alternative respiration pathway by AOX is also named as cyanide-resistant respiration (CRR). In recent years, the study of the alternative respiration pathway and alternative oxidase has been a hot topic in the area involving cellular respiration metabolism. In this review we summarized the latest research advances about the functions of alternative respiration pathway and alternative oxidase in industrial fungi.
Electron Transport ; Fermentation ; Fungi ; enzymology ; Industrial Microbiology ; Mitochondrial Proteins ; chemistry ; Oxidoreductases ; chemistry ; Plant Proteins ; chemistry

Objective To investigate the potential and early effect of hypertonic saline resuscitation on Tlymphocyte subpopulations in patients with traumatically hemorrhagic shock.Method Eighty-two patients with acute traumatically hemorrhagic shock admitted from Department of Emergency，Beijing Tongren Hospital，from De cember 2006 to July 2008 were randomly divided into：hypotonic saline(HS)group(n=43)and Lactated Ringer's solution(LR)group(n=39).The criteria of eligible patients were systolic blood pressure＜90mm Hg at admission with definite evidence of blood loss.Patients with immune system diseases and those who died within 24 hours of admission were excluded.Patients in HS group received intravenous administration of 200 mL of 7.5% sodium chloride withhin 15～20 minutes，and LR group received routine therapy with Lactated Ringer's solution.Blood pressure and heart rate were recorded 10，20，and 60 minutes after the start of resuscitation and compared between two groups.Before and 24 hours after treatment，peripheral blood levels of T-lymphocyte subpopulations including CD3+，CD4+，and CD8+ were measured by using direct immunofluorescence test and compared between two groups.The inter-group comparison was carried out by using independent sample t-lest and intragroup comparison using paired t-test.The numabers of operation，complication cases and death cases were conducted by using X2 test.SPAS version 11.0 software was used for statistical analysis.Results The volume of solution infused in HS group was(3820±623)mL and in LR group was(5430±1254)mL(P＜0.05).At 10 and 20 minutes after the sdministration of solution.the mean blood pressures in HS group were both significantly higher than those in LR group(P＜0.01).The levels of CD3+and CD4+lymphocytes in peripheral blood in HS group 24 hours after treatment were sinificantly higher than those in LR group(P＜0.01).Totally，63(76.8%)patients were cured and 19(23.2%)patients died.ARDS occurred in 10 patients and MODS occurred in 14 patiellts.The mortality，and the rates of ARDS and MODS in HS group were sinificantly higher than those in LP group(P＜0.05).Conclusiom In patients with acute traumatically hemorrhagic shock.HS can increase the effective circulating vol. ume，ameliorate the perfusion of tissues and organs，improve the immune fuction of T-lymphocytes，decrease the rats of ABDS and MODS，mad decrease the morality.HS is more effective than routine solution used for resuscitation.

BACKGROUND: Bushen Huogu decoction can effectively prevent and treat osteoporosis, but the concrete mechanism of pharmacology is still not clear. 25-hydroxy vitamin D3 and 1, 25-dihydroxyvitamin D3 are important coupling factors, which can regulate bone resorption and formation.OBJECTIVE: To investigate the curative effects of Bushen Huogu decoction on the bone mineral density, bone biomechanics, and level of 25-hydroxy vitamin D3 and 1, 25-dihydroxyvitamin D3 in blood serum, liver and kidney in the ovariectomized osteoporotic rats.METHODS: Totally 108 healthy female Sprague-Dawley rats were randomly divided into model group, sham-operated group, andtreatment group. All rats had been ovariectomized to induce estrogen absence and further establish osteoporotic models, except those in sham-operated group. Treatment group of rats were intragastrically administrated with 2 mL Bushen Huogu decoction,twice a day.RESULTS AND CONCLUSION: Compared with model group, the bone mineral density in the rat femur was significantly increased in the treatment group (P < 0.05), the index of maximal stress and maximal loading of the femoral head were also increased (P <0.05). The concentration of 25-hydroxy vitamin D3 and 1, 25-dihydroxyvitamin D3 in blood serum, liver and kidney were significant higher in the treatment group than those in the model group, and the levels were similar with those in sham-operated group (P >0.05). In the early period of estrogenic hormone absence, the Chinese kidney-tonifying drugs could activate bone metabolism, raise bone mineral density and reinforce quality of bone through up-regulating expression of 25-hydroxy vitamin D3 and 1,25-dihydroxyvitamin D3.

Objective To investigate the dates of 477 Chinese prenatal screening centers for previous half year analyse prena-tal screening status and provide recommendations for quality control.Methods All China prenatal screening centers were sent the data via the National Quality Assessment Scheme.This covered the software used,the risk cut-offs,monthly sample throughput,monthly median MoM of AFP,hCG,β-HCG,freeβ-HCG and uE3,monthly screening positive rate for trisomy 21,trisomy 18 and Open Neural Tube Defect.Results Screening protocol were versatile,with 73.48% (133/181)used two-marker model,24.31% (44/181)used three-marker model and 2.21% used four-marker model.About the software used, 350 laboratories never updated the screening parameters,89 laboratories had updated their median or parameter by manufac-turers,24 laboratories had updated the parameters by themselves.Cut-offs differed between laboratories.59.91% (275/459) used 1/270 as their cutoffs for trisomy 21.66.22% (296/447)used 1/350 as their cutoff for trisomy 18 and 96.52% (361/374)used cutoffs between 2.0~2.5 MOM for ONTD.Results of monthly median MOM:the percentage of laboratories that all six monthly median MOM within the target of 0.90~1.10 was 46.69% (155/332)for AFP,20.0% (4/20)for hCG, 29.17% (28/96)forβ-HCG,15.66% (31/198)for freeβ-HCG and 4.82% (11/228)for uE3.The percentage of laborato-ries that all six monthly median MOM within the target of 0.95~1.05 was 14.16% (47/332)for AFP,0% (0/20)for hCG,4.17% (4/96)forβ-HCG,12.63% (25/198)for freeβ-HCG and 4.82% (11/228)for uE3.About screening positive rate,there were difference of trisomy 2 1 positive rate in the same laboratory within the six month.Conclusion There were variations types of screening protocol,different kinds of soft platform,randomness of choice or update of medians or other important parameters and great difference in awareness of quality control.It would be taken effective and practical quality control measures to help laboratories improve quality control.

OBJECTIVE:To establish a method for the content determination of eudesmin in Zanthoxylum simulans. METH-ODS:HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-water(56:44,V/V)at flow rate of 1.0 ml/L,detection wavelength was 221 nm,column temperature was 25 ℃. RESULTS:The linear of eudesmin was 0.208-2.08 μg (r=0.999 8);RSDs of precision,reproducibility and stability tests were lower than 2.0%;recovery was 96.18%-103.04%(RSD=2.58,n=6). CONCLUSIONS:The method is simple and good reproducibility,and can be used for the content determination of eu-desmin in Z. simulans.

Background and purpose:As a new candidate tumor suppressor gene, p33ING1b has many biological functions. This study was done to investigate its role in the regulation of the proliferation of human colon cancer cell line SW480. Methods:The pcDNA3.1(+)/p33ING1b/SW480 cells were identified by Western blot and S-P immunohistochemical method. In order to elucidate the effect of expression of exogenous p33ING1b gene on the colorectal cancer cell SW480, the proliferation rates were analyzed by growth curves and colony formation assay in soft agar for cells including SW480, pcDNA3.1(+)/p33ING1b/SW480 and pcDNA3.1(+)/SW480. At same time, the apoptotic rate of cells and the cell cycle analysis were also tested by flow cytometry.Furthermore,using western blot analysis,we detected the expression level of the protein p53, p21WAF1,Bax and Bcl-2 in those three group cells, which initially indicate the molecule mechanism of inducing apoptosis by gene p33ING1b. Results:The cell growth rates of SW480 cells transfected with pcDNA3.1 (+)/p33ING1b were slower than those transfected with pcDNA3.1(+) or untransfected.The colony formation efficiency of pcDNA3.1(+)/p33ING1b/SW480 were decreased and the apoptotic rates were increased compared with pcDNA3.1(+)/SW480 and SW480(P0.05). Conclusion:After overexpression of exogenous p33ING1b protein in SW480 cell, there were inhibition of the proliferation rate and induction of apoptosis, the molecular mechanism might be associated with up- regulated expression of Bax and down-regulated expression of Bcl-2 by p33ING1b gene.

Objective To investigate the effect of pretreatment with U75302,antagonist of leukotriene B4 receptor 1 (BLT1),on cisplatin induced acute kidney injury in mice and its immunoregulatory mechanism.Methods Healthy C57BL/6 mice were randomized into four subgroups:1.healthy control group;2.cisplatin group;3.U75302 control group;4.cisplatin + U75302 group,n=6.Group 2 and 4 received intraperitoneal injection of cisplatin (20 mg/kg) on day 0,group 3 and 4received intraperitoneal injection of U75302 (5 μg/mouse) on day 0 and day 2.Mice were sacrificed on the 3rd day and blood and kidney were collected.Renal function and histological changes were estimated,the infiltration of immune cells were determined by flow cytometry,the level of peroxidase (MPO) in kidney were determined by colorimetry,relative expression of TNF-α,IL-1β,CXCL1,CXCL2 were detected by Real-time PCR.Results Compared with healthy control group,levels of BUN,Scr were higher in cisplatin group with serious tubular structural damage.There were more neutrophils,macrophages,CD4+ T lymphocytes,CD8+ T lymphocytes in kidneys of cisplatin group,the level of MPO and relative expression of TNF-α,IL-1β,CXCL1,CXCL2 were also higher in cisplatin group.Compared with cisplatin group,lower BUN [(17.75±1.80) mmol/L vs (42.6±6.66) mmol/L,P ＜0.05],Scr were found in cisplatin+ U75302 group with less tubular structural damage.Meanwhile,U75302 reduced infiltration of neutrophils [(146±13)×103/g vs (296±66) ×103/g,P ＜ 0.05],macrophages [(245± 13)× 103/g vs (420±78)× 103/g,P ＜ 0.05] in the kidney.Levels of MPO [(1.756±0.283) U/g vs (3.308±0.577) U/g,P＜0.05] and relative expression of TNF-α,IL-1β,CXCL1,CXCL2 were also lower.Conclusions BLT1 antagonist U75302 protects mice against AKI induced by cisplatin,and the mechanism is associated with reduced infiltration of inflammatory cells in kidney and the inhibition of kidney inflammation.

Objective To investigate the effect of lentivirus-mediated siRNA interference of USP39 on proliferation and migration of mice vascular smooth muscle cell in vitro.Methods Five siRNAs of siControl, siRNAUSP39-70, siRNAUSP39-71, siRNAUSP39-72 and siRNAUSP39-73 were designed and sythezied,mice VSMCs were infected with the lentivirus for delivering siRNAUSP39-73, and the stably transfected cells were selected by puromycin.The interference efficiency of siRNAUSP39-73 was assessed with Western blot.The effect of USP39 interference on the proliferation of VSMCs was determined by cells counting and MTT assay.Transwell assay was used to detect the migration of VSMCs.Results Recombinant lentiviral vector siRNAUSP39-73 was successfully transfected into mice VSMCs.Comparing with siControl group and Normal group, USP39 protein level of siRNAUSP39-73 VSMCs were decreased(P<0.05), and the proliferation and migration ability were all inhibited(P<0.05).Conclusion Targeted down-regulation of USP39 expression can inhibit the proliferation and migration of mice VSMCs in vitro.