BACKGROUND: Osteosarcoma cell OS-9607 is a cell line cultured from human osteosarcoma tissue. To construct cDNA library of this cell line so as to clone target gene from it.OBJECTIVE: To construct cDNA library of osteosarcoma cell OS-9607 by switching mechanism at 5' end of RNA transcript (SMART) technique and analyze its quality.DESIGN: Verified experimental study SETTING: Department of Orthopaedics, Changhai Hospital, Second Military Medical University of Chinese PLA MATERIALS: This experiment was carried out at the Department of Orthopaedics, Changhai Hospital, Second Military Medical University of Chinese PLA in May 2005. Osteosarcoma cell OS-9607 was cultured in CO2 incubator. Cells were pre-treated with DM for 30 minutes, recovered for 24 hours in standard culture medium. The whole RNA of osteosarcoma cell OS-9607 was isolated by RNAease reagent kit. The quantity and integrity of total RNA was detected by ultraviolet spectrometer and electrophoresis on a denaturing formaldehyde agarose gel stained by ethidium bromide (EtBr). METHODS: Total RNA was extracted from human osteosarcoma cell OS9607 and mRNA was isolated, cDNA of OS-9607 cells was acquired by in situ polymerase chain reaction (PCR) and long-distance PCR (LD-PCR),then the cDNA was ligated with dephosphorylated λ phage vector. The recombinants were constructed with SMART cDNA library construction kit.The size of cDNA inserts and the diversity of library were detected by PCR.MAIN OUTCOME MEASURES: The quality and integrity of total RNA, the titer of un-amplified and amplified library and recombination fraction; analysis of cDNA length of library RESULTS: ①The concentration of the isolated total RNA was determined and the measuring absorbance was between 260 nm and 280 nm. Its absorptance was about 1.9 A. 0.8％ denaturing formaldehyde agarose gel electrophorosis showed clear 28S rRNA and 18S rRNA,and the brightness rate of 28S rRNA and 18S rRNA (28S/18S) was 2:1. ②After linked to λ phage in vitro, packed and transfected host bacteria, the titer of unamplified library was 2.2×107 pfu/mL, and the titer of amplified 1 ibrary arrived to 4.5×1010 pfu/mL. The recombination fraction analysis showed that there were 8 blue plaques and 1 360 colorless plaques in one plate, so the recombination fraction was 99.5％. ③ The obtained cDNA library consisted of 1.6×106 recombinant bacteriophages and the average exogenous inserts of the recombinants was 1.5 kb.CONCLUSION: These results suggest that the obtained osteosarcoma cell OS-9607 cDNA library has high quality, which sets up a firm foundation for the further research on osteosarcoma cell OS-9607 and its elated genes screen.

This paper introduces the rapid & mobile first-aid unit from brigade or regiment medical aid station,especially discusses its equipment,performance,personnel structure,and working features.The rapid & mobile first-aid unit can perform emergency operation and anti-shock treatment for the serious wounded in frontier at any moment.It meets the requirements in such aspects as fast & effective first-aid,mobile & flexible treatment,high cure rate and good practicality.

Objective:To analysis the N-terminal amino acid sequence of 5D3Ag purified by mAb5D3 immunoaffinity column.Methods:SDS-PAGE and Western Blot were used to study the characteristics of 5D3Ag.From SDS-PAGE gel,5D3Ag was transferred to PVDF membrane and its N-terminal amino acid sequence was analysised with an auto amino acid sequencer.Results:SDS-PAGE of 5D3Ag showed a band on the gel with molecular weight of 43 kD.Western Blot study showed that mAb5D3 can recognize 5D3Ag.The N-terminal amino acid sequence was NQGSHGSEEQ.Conclusion:It is sure that 5D3Ag is unique to Osteosarcoma because of the purification method itself and the characteristics of mAb5D3.Compared with other reported Osteosarcoma associated antigen,5D3Ag is different from them in molecular weight and may be a novel Osteosarcoma associated antigen.With the help of N-teminal amino acid sequence,some primers may be designed to clone the whole length gene of 5D3Ag.The identification of 5D3Ag gene may be useful in understanding the biologic mechanics of Osteosarcoma and developing new diagnostic and therapeutic strategies.

Most patients with allergic conjunctivitis in children are vernal keratoconjunctivitis.Dry eye accompanied with al- lergic conjunctivitis in children is correlated with the decline of tear film stability.However,tear film stability decline is correlated with eosinophilic activation and concomitant release of inflammatory mediators.Moreover,conjunctival epithelial,goblet cells and tear film are damaged,then dry eye occur.Main symptom of dry eye in children is twinkling frequency increasing,followed by eye itch,eye red and photophobia.When allergic conjunctivitis in children is diagnosed,dry eye should be considered at the same time.Proper manage- ment for those allergic conjunctivitis can decrease ocular surface damages.

Adult ; Anemia, Aplastic ; chemically induced ; immunology ; therapy ; Benzene ; poisoning ; Cyclosporine ; therapeutic use ; Female ; Humans ; Immunity, Cellular ; Immunosuppressive Agents ; therapeutic use ; Male ; Middle Aged

Objective To discuss the value of body adiposity index( BAI) in the prediction of the risk of dysglycemia,dyslipidemia and hyperuricemia.Methods Five thousand and thirty residents as the participants from Jinshan New Area and nearby of Jinshan,Shanghai were enrolled,including 2004 males and 3026 females. The receiver operating characteristic curve( ROC) was drew to predict dysglycemia,dyslipidemia and hyperurice?mia by body mass index( BMI) ,waist circumference( WC) ,waist?hip ratio( WHR) and BAI in different gender groups.Medcale soft was used to compare the area under the curve.Results According to the ROC analysis,in males the area under curve in the prediction of dysglycemia of BMI,WC,WHR and BAI was 0.553,0.556,0. 538,0.540(P<0.05),and 0.513,0.523,0.523,0.535 in females(P<0.05).The area under curve in the predic?tion of dyslipidemia of BMI,WC,WHR and BAI was 0.641,0.626,0.563,0.588(P<0.05) in males and 0.617, 0.613,0.597,0.587(P<0.05) in females.The area under curve in the prediction of hyperuricemia of BMI,WC, WHR and BAI was 0.685,0.665,0.609,0.577(P<0.05) in males and 0.730,0.708,0.656,0.649(P<0.05) in females,and BMI, WC of AUC were higher than BAI both in males and females(P<0.05).Conclusion BAI could be a predictor for the risk of metabolic disease,but less than BMI and WC.

Objective To investigate the clinical significant and the difference of predicting the risk of dysglycemia and dyslipidemia by body mass index (BMI),waist circumference (WC) and waist-to-hip ratio (WHR) in order to look for the best predicting index.Methods Five thousand and thirty residents were participated to this study who were from Jinshan New Area and nearby of Jinshan,Shanghai,including 2004 males and 3026 females.They were divided into obesity group and non-obesity group based on the index of BMI,WC and WHR respectively.The ROC was made based on the above indies in different gender groups.Results The ROC area of BMI was the biggest with 0.641 (P ＜0.05) for male and 0.617(P ＜0.05) for female.The cut off value was 24.67 or 23.88 based on the male or the female.The same trend was seen in terms of WC with 88.5 cm for the male and 84.5 cm for the female.The small cut off value was seen in terms of WHR and there was no significant between male and female.Among male people,the cut off values was 26.01 in terms of BMI and 88.5 cm in terms of WC,0.89 cm in terms of WHR.Among the female people,there was the less predicting significant in terms of WC or WHR.The area under the curve had no significant differences,and the BMI predicted abnormal blood glucose of no value,the area under the curve is only 0.513 (P ＞ 0.05).Conclusion BMI is proved the best predictor for the risk of dyslipidemia.There are significant among BMI,WC and WHR in terms of predicting dysglycemia in different gender.BMI is proved without significant regarding of predicting dysglycemia in females.

Objective To generate finite element models of human head based on segmented computer tomography data.Methods A four-step procedure was adopted to configure the coarse mesh.The method of longest edge propagation path and the edge collapse were used to refine and optimize the final mesh.The method was evaluated by means of computer simulations in a 3-concentric-sphere head model and a three-layer realistic geometry human head model.Results The present simulation results showed reliability and rationality of the finite element computation,thus indicate the suitability of the developed method.Conclusion A multi-tissue finite element model is obtained by using this method.It can be applied to the computation of finite element based bio-mechanics and bio-electromagnetism.

Objective To explore the effect of ischemia-reperfusion injury on the retinal functions of rats. Methods Seventy Wistar rats were selected, 20 of which were selected randomly and divided into two groups (control group and single-irrigated group). The rats were anesthetized and their anterior chambers of the right eyes were cannulated with a 7-gauge needle connected to a reservoir containing ringers balanced salt solution, which was maintained at the same level of the eye for 1 hour. After that, ERG was recorded in both eyes of all rats. All the left rats were divided randomly into 10 groups and they were treated as the single-irrigated group. Retinal ischemia was induced by raising the reservoir to a height of 150 mm Hg. One hour later except the single ischemia group, all of the groups resumed perfusion after 3,6,12,and 24 hours and 3,5,7,14,and 21 days separately. ERG was recorded in both eyes of all rats. Results There was no difference in the results of ERG between left and right eyes in either the control group or the single-irrigated group. All the waves of ERG vanished in the single-ischemia group after 1 hour. In the ischemia-reperfusion groups, the waves of ERG partly recovered and the amplitude reduced persistently and progressively. Conclusion Ischemia-reperfusion injury may affect the function of the retina persistently and progressively.

AIM　To investigate protective effects and mechanism of tanshinones on ischemia-like injury models. METHODS　Six ischemia models including hypoxia, hypoglucose, oxidant injury, caffeine injury, nitric oxide neurotoxicity and excitatory amino acid injury were used to assay the anti-ischemic roles of tanshinones in cultured primary cortex neurons. The changes of injuried cortex neurons were observed by the way of morphological examination, and live neurons of crystal violet staining were measured according to absorbent index. RESULTS　It was found that tanshinones possessed obvious protective effects on primary neurons in injury models by the way of morphological examination. Crystal violet staining also indicated that tanshinones increased number of live neurons in injury models significantly. The protective effects of tanshinones on models of oxidant injury, caffeine injury and NMDA injury were superior to other injury models. CONCLUSIONS　83.0 μmol*L-1 tanshinones protected rat cortex cells from all injury models effectively in vitro.