Objective:To investigate the best preservation conditions of N-terminal pro-brain natriuretic peptide( NT-proBNP) and provide detection references with stable performance for detection kits.Methods: ELISA was used to quantitatively detect the changes in NT-proBNP contents in various preservation solutions.The effects of basic buffer system, preservative Proclin300 and antibiotics on the preservation of NT-proBNP were analyzed using univariate analysis.The combination of various factors was then optimized using orthogonal experiments, to identify the best preservation system for NT-proBNP.Results: The univariate analysis determined that the basic buffer system for NT-proBNP was 0.02 mol/L phosphate buffered saline(PBS) at pH7.2,the addition of pre-servative Proclin300 could extend the preservation time of NT-proBNP at 37℃ by one day, the combined addition of penicillin and streptomycin prolonged the preservation time of NT-proBNP by one day compared with individually adding penicillin or streptomycin.The orthogonal experiments identified a preservation solution for NT-proBNP as 20%calf serum,1/1 000 Proclin300,120 U/ml penicillin and 80 U/ml streptomycin in a basic buffer system of 0.02 mol/L PBS at pH7.2.This solution was used to preserve an NT-proBNP reference sample at 37℃.Seven days later,the calibrated fixed-value of the sample at 37℃was only 1.3%lower than that at 4℃.Conclusion:Optimized NT-proBNP serum preservation solution could preserve NT-proBNP standard sample at 37℃ for seven days.

Aim To make a research of rats with focal cerebral ischemia/reperfusion injury on pathological changes in brain and the changes of AQP4 and related proteins, in order to explore the relationship between AQP4 and brain edema. Methods Adult male SD rats, weighting 250~300 g, were randomly divided in-to Sham group and cerebral ischemia/reperfusion ( I/R) injury model group. The I/R model group was di-vided into the I/R-6 h, 12 h, 24 h, 48 h-four time point groups. The animal model of the right MCA is-chemia/reperfusion was established by suture method in mature SD rats. The nerve symptom score was con-ducted in the corresponding time points. Then, the permeability of brain tissue was detected by EB stai-ning;TTC staining was conducted to observe the cere-bral infarction volume;the dry wet weight method was used to detect the changes of brain water content; im-munohistochemical( IHC) , WB and RT-PCR were ap-plied to detect the expression of AQP4 , and the related factors at different time points of the model rats after is-chemia-reperfusion around infarcts. Results Com-pared with the Sham group, then ever function score of the rats in I/R model groups were much higher. With the increase of the reperfusion time, the cerebral in-farction volume, brain tissue permeability and the brain water content were also increased. IHC results showed that AQP4 expression gradually rose with widen distribution. WB and RT-PCR results verified the in-creasing level of AQP4 expression. The detection of the related proteins expression showed apparent changes. The expression of MMP-9 was increased, while the Oc-cludin and JAM-1 expression showed a decreasing trend. The I/R-48 h model group showed the most ob-vious differences in the expression of the related pro-teins and mRNA ( P <0. 01 vs Sham, respectively ) . Conclusion Accompanied with the aggravating cere-bral injury after cerebral ischemia/reperfusion, the ex-pression of AQP4 and MMP-9 level were activated, while the degradation of TJPs, Occludin and JAM-1, was increased. These factors are combined to make the formation of brain edema. This study makes a further research on the formation mechanism of the early stage for cerebral edema on I/R model and offers a potential for intervention in the filed of looking for a reliable drug therapy on cerebral edema.

Objective To analyse the effect of dual-antiplatelet drugs on S100β,IL-1β, adiponectin(ADPN)and NIHSS score in patients with acute cerebral infarction.Methods 58 patients diagnosed with acute cerebral infarction in first hospital of Qinhuangdao.All patients were collected and randomly divided into experimental group and control group according to random number table , 29 cases in each group.Both group were given the treatrnent of improvng the cerebral vascular circulation, protect brain cells, control blood pressure, blood glucose, oxygen when necessary.On the basis of conventional treatment, control group was treated with aspirin 200 mg, one time per day,orally.And experimental group was treated with clopidogrel 75 mg/d on the basis of control group,one time per day,orally.After treatment, the serum levels of S100β, IL-1β, ADPN and NIHSS score were detected in all patients.ResuIts After treatment, compared with control group,the serum S100βprotein level was significantly lower in the experimental group (P<0.05); the serum IL-1βlevel in experimental group was significantly lower (P<0.05);ADPN level in experimental group was significantly higher (P<0.05); NIHSS score of patients in experimental group was significantly lower (P<0.05).ConcIusions Dual antiplatelet drugs can reduce serum S100βprotein,IL-1βin serum of patients with acute cerebral infarction, increase the level of serum adiponectin, decrease NIHSS score, can effectively improre neurological function in patients with acute cerebral infarction.

Objective To analyse the effect of dual antiplatelet drugs on fibulin-5, vWF and P-selection in serum of patients with acute cerebral infarction.Methods Diagnosed 60 patients with acute cerebral infarction in our hospital and collected. All patients were randomly divided into experimental group and control group, 30 cases in each group.The control group was treated with conventional treatment and aspirin, and the experimental group was treated with clopidogrel on the basis of control group.After treatment, the serum levels of Fibulin-5, vWF, P-selection and adverse reactions were detected in all patients.ResuIts After treatment, compared with control group, the serum Fibulin-5 level was significantly lower in experimental group ( P<0.05 );the serum vWF level in experimental group was significantly lower ( P<0.05 );the serum P-selectin level in experimental group was significantly lower (P<0.05);there was no significant difference in the incidence of adverse reactions between two groups. ConcIusion Dual antiplatelet drugs can reduce serum vWF, P-selectin and fibulin-5 in serum of patients with acute cerebral infarction, adverse reactions do not significantly increase, have guiding significance to clinical application.

Objective To study the effect of bel-2 siRNA on apoptosis of HL-60 cells.Methods bcl-2 siRNA was synthesized in vitro transcription with silencer siRNA construction kit.The synthesized siRNA was transfected into HL-60 cells with Amine siPORT transfection.We used MTT flow cytometer and hoechst 33258 flourescence stainning t0 evaluate cell proliferation and apoptosis. Results.Bcl-2 siRNA could partially inhibit the growth of HL-60 cells.After incubated with bcl-2 siRNAl for 48 hours,HL-60 cells exhibited morphologic characteristic of apoptosis including chromatin condensation,crescents formation and nuclear fragmentation.Conclusion Effective bcl-2 siRNA can induce apoptosis and inhibit cell proliferation.

Objective To investigate the effects of cryopreservation on the growth and osteogenesis capa- bility of human bone marrow stromal cells(BMSCs)on demineralized bene matrix(DBM).Methods Bone marrow aspirates were obtained from the lilac crests of three donors.The BMSCs were isolated from the bone marrow by density gradient centrifugation.Cells of passage 3 were cryopreserved in liquid nitrogen for 24 hours,and then re- covered.The non-cryopreserved BMSCs were used as the control,The cryopreserved and control BMSCs were cul- tured in osteogenic media,collected and labeled with Dil to be seeded onto the DBM when cells were confluent.The percentage of BMSCs adhered to the DMB was detected.The cell morphology and matrices secreted by BMSCs on the DBM were observed by the inverted phase-contrasted microscope,fluorescence microscope and scanning electron microscope(SEM).The growth and viability of BMSCs on the DBM were determined using the modified MTT ashy. The osteogenesis ability of BMSCs on the DBM was determined by assessment of the alkaline phosphatase(ALP) activity and osteocalcin(OCN)content.Results The percentages of the cryopreserved and control cells adhered to DBM were(97.25?1.17)% and(97.00?1.09)% respectively.The cells adhered well to the DBM and grew rapidly.Large amounts of matrices on the DBM were observed by the light microscope and SEM.The cells embedded in the matrices could be observed by fluorescence microscope.There were no significant differences in the assay values of MTT,ALP and OCN between the cryopreserved and control BMSCs on the DBM.Conclusion Since cryopreservation does not affect the growth and osteogenesis capability of BMSCs on DBM,the cryopreserved BMSCs can be used as a cell source in bone tissue engineering.

Objective To explore the value of dual-source CT spectrum energy curve and iodine quantification in determining the degree of differentiation of clear cell renal carcinoma(ccRCC).Methods The radiographic data of 38 cases with pathologically confirmed ccRCC were analyzed retrospectively.All patients underwent dual-source CT,cortical phase and nephrographic phase dual energy scan,including 22 well-moderated differentiated(gradeⅠ,gradeⅡ)and 16 poorly differentiated neoplasms(gradeⅢ,gradeⅣ).The slope of spectrum energy curve (40-70 keV,70-100 keV,100-140 keV)and the normalized iodine concentration (NIC)between the two grades were compared. Results In the spectrum energy curve 40-70 keV and 70-100 keV segment interval,the slope of spectrum of well-moderated differentiated group was higher than that of poorly differentiated group,the difference was statistically significant(P<0.05).In the 100-140 keV segment interval,the slope of spectrum of well-moderated differentiated group was higher than that of poorly differentiated group, but the difference was insignificant(P>0.05).The NIC of well-moderated differentiated group was significantly higher than that of poorly differentiated group(P<0.05).Conclusion The slope of spectrum energy curve and NIC are correlated with the differentiation degree of ccRCC.The slope of spectrum energy curve and iodine quantification of dual-source CT could be used as new methods for preoperative grading of ccRCC.

OBJECTIVE: To explore the management of the laryngeal injury. METHOD: The forty five patients of laryngeal injury had been rescued. Twenty eight patients were treated with the tracheotomy and twenty four patients of them were treated with the laryngoplasty. RESULT: All the patients were survived. Tracheostomy tube of twenty eight patients was taken away without the cicatricial structure of larynx. CONCLUSION With the reasonable management after the laryngeal injury, the injury could be cured in time without complications.
Adolescent ; Adult ; Aged ; Female ; Humans ; Larynx ; injuries ; Male ; Middle Aged ; Neck Injuries ; surgery ; Young Adult

BACKGROUNDTo evaluate therapeutic outcome of limited stage small cell lung cancer treated with chemotherapy alone or combined with radiation therapy with different doses.

METHODSA retrospective analysis was performed in 128 limited-stage small cell lung cancer patients who were treated with three different ways of treatment, from February 1988 to March 1998 in Heilongjiang Cancer Hospital. All patients were pathologically proved. Forty-two patients received chemotherapy alone (C), 48 patients were treated by interdigitating chemoradiotherapy (IDG) and other 38 patients received concurrent chemoradiotherapy (CON). For thoracic radiation, 20 patients received a dose of ≤45 Gy, 23 ≥60 Gy, and 43 > 45 Gy but < 60 Gy .

RESULTSThe 3-year survival rates were 23.7%, 20.8% and 4.8% in the CON, the IDG and the C groups respectively. There was a significant difference between the CON and the C groups ( P < 0.05), as well as between the IDG and the C groups ( P < 0.05). There was no remarkable difference between the CON and the IDG groups ( P > 0.05). Loco-regional recurrence rate was significantly higher in ≤45 Gy group (55.0%) than that in ≥60 Gy group (8.7%)( P < 0.01).

CONCLUSIONSThe chemotherapy combined radiotherapy may improve the survival of patients with limited-stage small cell lung cancer. Dose of thoracic radiation might be related to the loco-regional recurrence.

Aim To study the apoptosis effect of cucurbitacin Ⅱa on non-small cell lung cancer cell lines NCI-H460 and A549 and its underlying mechanism.Methods Cell viability was assessed by CCK-8 assay.The apoptosis effect and cell cycle arrest were detected by Flow cytometry.Western blot was employed to detect the related protein.Results The proliferation of lung cancer cell lines NCI-H460 and A549 was inhibited by CuⅡa, which showed cytotoxic activity with IC50 values of 224.9 nmol·L-1 and 108.3 nmol·L-1 against NCI-H460 and A549 respectively.CuⅡa induced the cells apoptosis and cell cycle arrest at G2/M phase.The results of Western blot showed CuⅡa inhibited the phosphorylation of STAT3 and Cofilin in a dose-dependent manner.Further, CuⅡa inhibited the phosphorylation of Aurora A, in line with the important characteristics of anti-tumor effect of Aurora A kinase inhibitor with blocking cells in the G2/M phase.Conclusion CuⅡa has obvious anti-tumor effect against non-small cell lung cancer, which suggests its value as a lead compound for lung cell carcinoma.