Objective To establish a multiplex suspension array for simultaneously detecting five species of important arbovirus: dengue virus (including four serotypes), japanese B encephalitis virus, west nile virus, tick-borne encephalitis virus and yellow fever virus. Method All published genomic sequences of the above arboviruses were collected from GeneBank, and then, the genus universal primers and specific probes were designed according to the conservative and specific NS5 region of the genus Flavivirus. The amine-modified probes were covalently coupled to different carboxylated microspheres by carbodiimide methods. By combining general RT-PCR with virus-specific probe hybridization, a Luminex xMAP system-based suspension array, which could simultaneously differentiate five arboviruses and discriminate 1～4 serotypes of dengue virus, was successfully established. Results A cutoff value twice higher than that of the background fluorescence was judged as positive reaction. RT-PCR products of many arboviruses were used to evaluate the specificity and repeatability of the array, the results showed that the assay was highly specific and had good repeatability. Meanwhile, the coefficient of variability was less than 8% from inter- and intra- assay, implying that both the repeatability and stability of the array were good. Furthermore, several viruses which had been quantitated by plaque assay were used to determine the array’s sensitivity. The results showed that the array’s detection limits were about 1.4 PFU for JBEV and YFV, 14 PFU for WNV respectively. 55 clinical and cultured samples were detected by the established assay, 16 out of the 55 samples were confirmed to be positive. There was only one sample which was not detected contrasted with conventional RT-PCR. However, it had advantages in detecting complex samples rapidly. Conclusion A multiplex suspension array for simultaneously detecting and serotyping five species of important arboviruses has been successfully developed, and it may play an important role in diseases diagnosis and prevention as well as epidemiological studies.

Multiple tumor makers are needed to improve the diagnostic rate of the simultaneously detection of malignant tumors through screening. Therefore, multiplex detection technology is urgently required to improve the screening efficiency. Suspension arrays are multiplex detection method based on gene microarrays. It consists of encoded microbeads, probes, targets, and report molecules are applied to analyze targets quantitatively. The microbead encoding strategy is a hotspot in suspension array research. The photonic crystal encoding mentioned in this review is a type of optical encoding that is very stable and easily decoded. Photonic suspension arrays have broad prospects in the screening and diagnosis of malignant tumors through long-term studies. This review summarizes the basic principle, classification, and characteristics of photonic suspension arrays and their application in the screening of malignant tumors.

Objective To develop a diagnostic test based on indirect immunofluorescence assay(IFA) to detect special antibodies in the serum of SARS patients, thus to provide a reference material for confirmation of the clinical diagnosis of SARS. Methods SARS coronavirus GZ01 and BJ01 strains isolated in our laboratory were used to infect Vero E6 cells. When CPE reached 25%, cells were trypsinized and transferred to 10 well slides in a quantity of 40?l with a cell density of 2?10 7 /ml. After 4 hour incubation at 37℃,the slides were fixed with acetone, and IFA was used to detect antibodies in serum samples, which were obtained from 154 SARS patients and 14 non SARS patients with respiratory disease, as well as 100 healthy volunteers. Results IFA method for detecting antibodies of SARS coronavirus was developed. Sera from one hundred and forty two out of 154 clinically diagnosed patients were IFA positive, with a positive rate of 92 3%. Sera from 14 non SARS patients with respiratory disease and 100 healthy persons were all IFA negative. Conclusion The IFA method we developed was sensitive and specific in detecting SARS antibodies in serum, and was a reliable test for laboratory diagnosis of SARS coronavirus.

BACKGROUND: Patients who suffered total hip replacement are mostconcerned about the survivorship of prosthesis. OBJECTIVE: To evaluate the postoperative curative effect following ce mented and cementless THR with a retrospective method, so as to provideexperience for prolonging the survivoship of prosthesis. DESIGN: Randomized and controlled observation. SETTING: General Center of Orthopaedic Department, General Instituteof Bone Oncology, Tangdu Hospital, Fourth Military Medical University ofChinese PLA. PARTICIPANTS: We admitted 356 patients who underwent THR fromDepartment of Orthopaedics, Tangdu Hospital, Fourth Military Medical U niversity of Chinese PLA between March 1993 and March 2004. Amongthem, 298 were contacted and 105 (108 hips) followed up. The patientsparticipated in the review voluntarily. They were of either gender and haddifferent types of prosthesis. Prosthesis made in China was adopted before2000 and prosthesis made in American STRIKER company after 2000: Prosthesis made in China was made of home-made bone cement; Prosthesisbone cement (import) was provided by American STRIKER prosthesiscompany. Home-made bone cement and import have the same components. Barium was added in both bone cement . The whole operation was con ducted by the physicians who worked in the artificial joint department afterexamination. METHODS: According to informal discussion summary about total hipreplacement of Chinese Journal of Surgery in 1982 and Evaluation Scale ofMayo Total Hip Replacement Curative Effect, we designed follow-up tableby ourselves. Totally 105 (108 hips) patients were followed up, amongthem, 62 (63 hips) were in the cemented group, 43 (45 hips) in the ce mentless group. Pain, function and motion range of the patients and X-raywere evaluated and analyzed respectively. MAIN OUTCOME MEASURES: ① Postoperative pain degrees. ② Postoperative function of hips. ③ Postoperative motion range. ④ width oflight around the prothesis , distance of horizontal or vertical shift of theprosthesis. ⑤ range of ectopic ossification of the prosthesis. ⑥Osteolysisdegree of proximal femur. RESULTS: ①There was no significant difference of lateral femoral painduring follow-up period [Cemented group: 24 hips (38.5%) ,cementlessgroup: 18 hips(40.0% )]. ② Limping appeared in the both two groups ③ There was no significant difference of range of motion above 160° betweentwo groups (Cemented group: 62 hips; cementless group: 44 hips). ④Therewas no significant difference in subsidence of femoral prosthesis and hori zontal or vertical shift of acetabular prosthesis between two groups . ⑤ There was no significant difference of re lative value of femoral proximalbone density between cemented group [57.4(9-118)] and cementless group[72.8( 14-130)]. ⑥There was no significant difference of postoperative cu rative effect, possible survival rate of prosthesis and femoral proximal ex tensive osteolysis of the patients between the two groups. CONCLUSION: Postoperative curative effect of the patients between ce mented group and cementless group are similar, both not obtaining an idealfixed effect. The choice of prosthesis type does not affect the survivorship of prosthesis, but it depends on the age of patients to decide whether rebuilding is necessary or not: Osteolysis is not related to age, gender or prosthesis type of the patients.

Objective To introduce a new design of interlocking system which is easy in use, makes the distal fracture fragments well aligned and rigidly fixated, reduces exposure to radiation for the surgeon and patient, and allows for a decrease in operating time. Methods The new interlocking system was designed to be a self-locking nail with bevels (b-SLN). According to the bevel principle, the axial force of the nail should be turned into a transverse force so as to control the transverse nail's movement. 10 dry-bone simples were made into the fracture model of the middle-femur to test the prototype nail. The retention of the nail in the intramedullary cavity was evaluated by the radiogram. The mechanical properties of a simulated model of single leg loading were tested on the instron-1342 type MTS. Results The prototype nail performed well as designed. The process was stable and reliable. The retention in the intramedullary cavity was fine. The shearing force between the transverse nail and main nail was enough to cut anything around them. The mechanical properties of b-SLN were similar to those of the Grosse-Kempf nail, better than auto fork compress locked intramedullary nail, and much better than MHUA nail and Ender's nail. Conclusions The structure of b-SLN is simple and reliable. There is no focus-point under stress. Its biomechanic properties are satisfactory. It is easy to use with no need of fluoroscopes in the operating theatre.

Objective To observe the clinical efficacy of alprostadil combined with valsartan in treatment of chronic glomerulonephritis proteinuria.Methods 78 patients with chronic glomerulonephritis proteinuria were randomly divided into two groups,and each group had 39 cases.The control group was given conventional treatment,while the observation group was given alprostadil combined with valsartan on the basis of the control group.The clinical outcomes were compared between the two groups.Results The total effective rate of the observation group was 92.31％,which was significantly higher than 74.36％ of the control group (x2 =9.825,P ＜ 0.05).After treatment,the 24h Upro,BUN and SCr of the observation group were (1.00 ± 0.39) g/24h,(7.11 ± 0.15) mmol/L and (80.86 ± 0.65) μmol/L,which were significantly lower than those of the control group [(1.30 ± 0.48) g/24h、(9.18 ± 2.21) mmol/L and (98.71 ± 4.34) μmol/L],the differences were statistically significant (t =9.32,7.83,7.12,all P ＜ 0.05).Conclusion Alprostadil combined with valsartan in the treatment of chronic glomerulonephritis proteinuria has significant effect,and it can significantly alleviate clinical symptoms,improve renal function,which should be widely applied in clinical.

BACKGROUND: At present, there are only morphological observation about the femoral osteolysis in domestic literatures and reports of quantita tive study are also few.OBJECTIVE: To quantitatively analyze the changing status of osteolysis in proximal femur after total joint replacements (TJR) through the changing rules of bone density in proximal end of femur.DESIGN: Observation on patients before and after self-comparison.SETTING: Department of Orthopedics, Tangdu Hospital of Fourth Military Medical University of Chinese PLA.PARTICIPANTS: Twenty-eight hips in 26 patients received total hip replacements in the Department of Orthopedics, Tangdu Hospital of Fourth Military University from March 1994 to March 2004, including 16 hips in 14 male patients and 12 hips in 12 female patients, were selected. Types of prostheses: 17 hips in 16 patients with ameliorated Moore cemented prostheses were taken as cemented prosthesis group with a mean age of 57 years; 11 hips in 10 patients with microbores on the surface of implant were taken as cementless prosthesis group with a mean age of 55 years.Patients were diagnosed as traumatic arthritis before operation with the mean Harris score of 49 (20-77) points on hip. Age, gender and diagnosis after operation in patients of two groups were matched.METHODS: The gray degrees of X-ray in patients of two groups were measured with computer image analyzing, relative values were obtained,I.e. The average gray degrees of 1.0-2.0 cm2 area in femoral trochanter and iliac bone were measured with the same X-ray; Differences between them represented the relative value of bone density in greater trochanter of femur. Changing rules of bone density were quantitatively analyzed.MAIN OUTCOME MEASURES: Relative values of bone density in greater trochanter of femur of patients in two groups one week before and after operation, during the follow-up time were measured respectively.RESULTS: A total of 26 patients (28 hips) with the follow-up time of 9-126months were all involved in the analysis of results. Relative values of bone density were increased to different degree one week after operation. And rate of osteolysis was 100% with a mean value of 57.4 (9-118) in greater trochanter of femur from 17 hips of 16 patients in the cemented group during the follow-up time; Relative value of bone density in greater trochanter of femur in 10 hips of 11 patients in the cementless group during the follow-up time were increased to different degree one week after operation.Rate of osteolysis changing was 100% with a mean value of 72.8 (14-130).Differences between two groups were not significant (P ＞ 0.05). Changes of osteolysis firstly appeared in the 9th month of post-operation and were obvious 2-4 years after operation and lessened in the 6th year.CONCLUSION: There are osteolysis in cemented and cementless prostheses after total hip replacement and no significant differences between two kinds of prostheses.

Objective:To study immunosuppression mediated by the porcine FcγRⅢ in porcine reproductive and respiratory syndrome virus ( PRRSV ) infection to pulmonary alveolar macrophages ( PAMS ).Methods: In this study pulmonary alveolar macrophages cells were treated with containing 200 TCID50 PRRSV,lipopolysaccharide (LPS) (100 ng/ml) and purified mouse anti-pig FcγRⅢIgG (550 μg/ml) separately,simultaneously,PAM cells treated with purified mouse anti-pig FcγRⅢIgG (550 μg/ml) was infected by 200 TCID50 PRRSV ,untreated PAM cells as the control group.Each group were post-cultured 12,24,36,48,60,72 h, the cells and the supernatant were collected.The dynamic variation of PRRSV RNA copies in inoculation group were detected by using real-time fluorescence quantitative PCR method.mRNA level of IFN-αand TNF-αin each group were detected by using relative fluorescence quantitative PCR.Results:The result showed that mRNA level of IFN-αwas improved during PRRSV infection to PAMS 12-24 h,and mRNA level of IFN-αwas inhibited during 36-72 h,then mRNA level of IFN-αrecovered normally; mRNA level of TNF-αwas increased slightly post-infection 12-72 h.IFN-αand TNF-αmRNA levels of PAM cells treated with LPS were both up-regu-lated,using the purified mouse anti-pig FcγRⅢ IgG to treat the PAM cells,selective activation of porcine FcγRⅢ in the PAM cells down-regulated significantly mRNA levels of IFN-αand TNF-α.PRRSV infection assay mediated by selective activation FcγRⅢof the PAM cells inhibited antiviral cytokine ( IFN-αand TNF-α) mRNA levels.Conclusion:The results show selective activation of FcγRⅢinhibited significantly mRNA levels of the antiviral cytokine IFN-αand TNF-αof host cells,and innate antiviral immune response to PRRSV infection.

Objective To identify the specific microRNA (miRNA) that can be taken as a molecular marker for human bone marrow-derived mesenchymal stem cells (MSCs). Methods MSCs were isolated and cultured from bone marrow through density centrifugation and then were induced to differentiate into osteoblasts and chondrocytes. Samples of MSCs, osteoblasts and chondrocytes were detected by miRNA microarrays single channel fluorescence chip to determine the expression levels of miRNAs. Significance Analysis of Microarrays ( SAM, version 2.1 ) software was used to analyze the raw data to determine the miRNAs overexpressed in MSCs, which was validated in the same sample using real time reserve transcriptase polymerase chain reaction (RT-PCR). Results MSCs were successfully isolated from bone marrow and induced to differentiate into osteoblasts and chondrocytes in vitro. Microarrays showed that eight miRNAs (has-miR-424, has-miR-34a, has-miR-593, has-miR-10a, has-miR-148a,has-miR-602, mmu-miR-709 and mmu-miR-665) were overexpressed in MSCs but underexpressed in osteoblasts. Three miRNAs including has-miR-424, PREDICTED_MIR189 and mmu-miR-665, were overexpressed in MSCs but underexpressed in chondrocytes. The has-miR-424 expression in MSCs was 6.6times higher than in osteoblasts and 4.4 times higher than in chondrocytes. The results of real time RTPCR showed that the miR-424 was overexpressed in MSCs, 3.6 times higher than that in osteoblasts and 3.1 times higher than that in chondrocytes, which was coincident with the results of microarray. Conclusions The screened MSCs express more miRNAs in comparison with osteoblasts and chondrocytes,play important roles in maintaining self renewal and undifferentiation of MSCs and is a promising specific molecule marker for MSCs.

Background and objectives Hyperhomocysteinemia is an independent risk factor for cardiovascular disease. Homocysteine thiolactone (HcyT), one of the homocysteine metabolites in vivo, is toxic both in vivo and in vitro. The aim of this study was to investigate the effect of HcyT on apoptotic damage in human umbilical vein endothelial cells (HUVECs) and the role of antioxidants in the reduction of HcyT-induced apoptosis. Methods HUVECs were cultured in DMEM supplemented with 20% heat inactivated fetal bovine serum cell cultures were maintained in a humidified 5% CO2 atmosphere at 37 ℃. Cytotoxicity was determined by MTT assay,which consists of hypodiploid cells with propidium iodide labeling and intracellular reactive oxygen species levels using 2',7'-dichlorofluorescein diacetate as the probe by flow cytometry. Results HcyT (250-2000μM) induced HUVECs apoptosis in a time- and concentration-dependent manner. Reactive oxygen species levels rose in response to increasing HcyT concentrations at 24-h incubation.The reduction of cell apoptosis by N-acetylcysteine, vitamin E, or pyrrolidine dithiocarbamate, occurred simultaneously with a significant decrease in intracellular reactive oxygen species levels. Conclusion HcyT exerts its cytotoxic effects on endothelial cells through an apoptotic mechanism involving cellular reactive oxygen species production. The capacity of N-acetylcysteine, vitamin E, and pyrrolidine dithiocarbamate to scavenge HcyT-induced cellular reactive oxygen species correlates well with their efficiency to protect against HcyT-promoted apoptotic damage. The protective effect of pyrrolidine dithiocarbamate on cell apoptosis indicates HcyT-generated hydrogen peroxide may provoke cell apoptosis via activating nuclear factor-kappa binding protein.