Objective Present study aimed to research the mechanism of the use of 0.2% brimonidine tartrate eye drops in the treatment of optic nerve crush injury of rat.MethodsAnimal models of optic nerve crush injury were created in 60 SD female rats by clipping the exposed optic nerve at 2 mm in retrobulbar for 6 seconds with 78 grams of reverse forceps.The successful model was identified as Marcus-gun pupil without bleeding of fundus after operation.The animals were randomly assigned to model group and brimonidine treating group,and another 30 normal SD rats were used as the normal control group.The 0.2% brimonidine tartrate eye drops was topically administered at 2 hours before operation and after operation twice per day in the brimonidine treating group.The retinas from 18 rats were isolated after 3,7,21 days for RGCs counting by H&E staining,and the retinal ultrastructure was examined under the transmission electron microscope.The retinas from the other 72 SD rats (including normal,model and brimonidine groups) were prepared for the detection of bcl-2 and bax using immunohistochemistry l,3,5,7,14,21 days after the operation.ResultsNormal and almost normal retina structure was exhibited in rats of the normal group and brimonidine treating group,but disorder of cellular arragement and decrease of retinal thickness were found in the model rats under the optical microscope.The RGCs counting was significantly different among the three groups from 3 days through 21 days after operation with the considerably declination in the model group and brimonidine treating group compared with the normal control group (P＜0.05-0.01).However,that of the brimonidine treating group was obviously increased in comparison with the model group (P＜0.01).The expression of bax in rat retina was obviously reduced (P＜0.01),but the expression of bcl-2 was increased in brimonidine group compared with the model group from 5 through 7 days after operation (P＜0.01).ConclusionThe 0.2% brimonidine tartrate eye drops has a preventive effect on optic nerve crush injury of rat,and its inhibition on apoptosis is one of the mechanisms.

In order to construct and express human cardiac troponin C-linker-troponin I(P) [ cTnC-linker-TnI(P)] fusion protein, detect its activity and prepare lyophilized protein, we searched the CDs of human cTnC and cTnI from GenBank, synthesized cTnC and cTnI(30-110aa) into cloning vector by a short DNA sequence coding for 15 neutral amino acid residues. pCold I-cTnC-linker-TnI(P) was constructed and transformed into E. coli BL21(DE3). Then, cTnC-linker-TnI(P) fusion protein was induced by isopropyl-β-D-thiogalactopyranoside (IPTG). Soluable expression of cTnC-linker-TnI(P) in prokaryotic system was successfully obtained. The fusion protein was purified by Ni²⁺ Sepharose 6 Fast Flow affinity chromatography with over 95% purity and prepared into lyophilized protein. The activity of purified cTnC-linker-TnI(P) and its lyophilized protein were detected by Wondfo Finecare™ cTnI Test. Lyophilized protein of cTnC-linker-TnI(P) was stable for 10 or more days at 37 °C and 4 or more months at 25 °C and 4 °C. The expression system established in this research is feasible and efficient. Lyophilized protein is stable enough to be provided as biological raw materials for further research.
Escherichia coli ; Freeze Drying ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; Troponin C ; biosynthesis ; Troponin I ; biosynthesis

Objeictivie To investigate the diagnostic value of anti-mutated citrullinated vimentin (anti-MCV) antibody, anti-cyclic citrullinated peptide (anti-CCP) antibodies and glucose-6-phospha-teisomerase (GP1) for rheumatoid arthritis (RA). Methods Anti-MCV antibody, GPI and anti-CCP antibody were detected in serum samples of 109 RA patients, 24 non-RA rheumatic diseases patients and 19 healthy blood donors. The sensitivity and specificity of these parameters for the diagnosis of RA were analyzed. Results Both the positive rate and average cut off concentration of anti-MCV and GPI in RA were higher than those of non-RA rheumatic diseases or healthy controls (P<0.05). A significant difference was found between anti-MCV and GPI in RA patients. The most sensitive and specific parameter in RA was anti-MCV (99.1%) and anti-CCP (90.7%) respectively, but, when anti-MCV combined with anti-CCP, or GPI or anti-CCP and GPI, the specificity could be up to 98.1%. Coniclusions Anti-MCV, anti-CCP and GPI alone or in combination may be valuable parameters for the diagnosis of RA.

OBJECTIVE： To study the method of measuring the in vitro dissolubility of lansoprazole enteric－ coated tablets and to find out the difference between the dissolubility of lansoprazole enteric－ coated tablets and that of lansoprazole enteric－ coated capsules.METHODS： To establish a measuring method of the dissolubility of lansoprazole enteric－ coated tablets and compare the dissolubility of lansoprazole enteric－ coated tablets with that of lansoprazole enteric－ coated capsules and to acquire the dissolving parameters for variance analysis.RESULTS： (1)The accumulative dissolving quantity of the both preparations exceeded 80％ in 30 minutes.(2)The comparison of Weibull parameters,T50,Td and m,between each other in group A showed no significant differences(P>0.05).It was the same for those of group B.(3)The parameters,T50 and Td,of group A,in comparison with those of group B,showed significant differences(P<0.001).The times needed for dissoiving 50％ and 63.2％ of quantity in group B were both shorter than those in group A.The m parameters in different groups had no significant differences(P>0.05).There was no significant difference in slope of equation between two groups.CONCLUSION： This measuring method of the in vitro dissolubility of lansoprazole enteric－ coated tablets is simple,convenient,accurate,stable,feasible and good in repetition.It is concluded that there is significant difference between the dissolubility of the two dosage forms of enteric－ coated preparation of lansoprazole.

Objective To investigate the apoptosis mechanism induced by BH3 mimetic S1 in human leukemia cell line K562.Methods Cell viability was detected by XTT to S1 in leukemia cell line K562.K562 cells was incubated with S1 for different time,the apoptosis rate of K562 cells was determined by flow cytometry analysis.Caspase-3,-8,and-9 activities were measured by absorption spectra.Co-immunoprecipitation was used to analyze the releasing of bax,bak from bcl-2 and mcl-1.Results Compared with control group,a dose-dependent increase in apoptosis coincided with a dose-dependent decrease in cell viability following S1 treatment suggested that S1 inhibits cell proliferation through the induction of apoptosis.The IC50 value at 24 h for S1 was 13.5 μ mol/L.Exposure of K562 cells to S1 for 12 h resulted in a time-dependent increase in FITC-Annexin-positive/PI-negative early apoptotic cells.The strong increase of FITC Annexin/PI doublepositive cells after a 24 h treatment indicated a shift to late apoptosis.S1 activated Caspase-3 and-9,but not Caspase-8 indicated that S1 induced K562 cells apoptosis via the intrinsic pathway.K562 cells treated with 5 μmol/L of S1 showed a disruption in bcl-2/bax,mcl-1/bak complexes after 8 h S1 treatment.Conclusion The main mechanism that S1 induces K562 cells apoptosis might be through the inhibition of bcl-2/bax,mcl-1/bak complexes dissociation.

Objective To observe the effects of high sucrose,high saturated fatty acid and high unsaturated fatty acid(diets) on insulin resistance and endothelium-dependent vasodilation function.Methods Adult Wistar rats were divided into normal control(NC) group,high sucrose(HS) group and high saturated fatty acid(HSF) group,high unsaturated fatty acid(HUF) groups.Insulin sensitivity was tested by hyperinsulinemic-euglucemic clamp after 24 weeks.Acetylcholine-induced(or sodium nitroprussideinduced) relaxation of preconstricted isolated renal arteries was measured by Mulvany myograph.Results GIR was obviously lower in experimental groups than that in NC group.GIR was negatively correlated with triglyceride (TG),free fatty acid(FFA).Acetylcholine-induced relaxation was markedly decreased in all experimental groups compared with that in NC group and the maximal response was decreased 37.4% in HSF group,32.7% in HUF group,27.7% in HS group.Acetylcholine-induced relaxation was enhanced by incubation with L-Arg and decreased incubated with L-NNA,MB in all experimental groups.Vasodilation response was negatively correlated with TG,INS and well positively correlated with NO,GIR.There was significantly negative correlation between FFA andNO.Conclusions: The rats fed high sucrose,high saturated fatty acid and high unsaturated fatty acid diets developed insulin resistance with reduced endothelium-dependent vasodilation function.

OBJECTIVE To strengthen management of nosocomial infections in comprehensive intensive care unit so as to enhance medical treatment quality.METHODS Nosocomial infection administration department constituted the management institutions and standards of nosocomial infections with supervising,implementing targets monitoring.Comprehensive intensive care unit instituted several groups of nosocomial infection management and fulfiled the precautions and control measures.RESULTS By means of interventions and prophylaxes,the occurrence of nosocomial infection in comprehensive intensive care unit decreased.CONCLUSIONS It ensures for medical treatment quality and safe by strengthening the management of nosocomial infections.

Objective To investigate the ability of single photon emission computed tomography (SPECT) and MRI in detecting skull-base invasion in nasopharyngeal carcinoma. Methods Sixty-one patients with nasopharyngeal carcinoma received whole body and skull-base tomography SPECT, and nasopharynx and skull-base MRI before radiotherapy. The results were double-blind compared and evaluated. Results The overall positive rates of skull-base invasion detected by SPECT and MRI were 51% and 46% (P=0.508). In paitents with headache, cranial nerve palsy or both, the rates were 83% and 86% (P=1.000) ,80% and 80% (P=1.000), 88% and 94% (P=1.000), respectively. In patients with T1+T2 and T3+T4lesions,the rates were 22% and 0(P=0.031) ,74% and 82% (P=0.250) ,repectively. In patients with N0+N1and N2+N3lesions,they were 50% and 48% (P=1.000) ,53% and 40% (P=0.500) ,respectively. The conformation rate between SPECT and MRI was 85%. Binary Logistic regression analysis showed that T stage was a risk factor for positive SPECT(χ2=4.23,P=0.040, OR=3.04). Headache tended to be a risk factor for both positive SPECT and positive MRI (χ2=3.13, P=0.077, OR=4.54;χ2=3.64,P=0.056,OR=12.00). Conclusions The detection sensitivity of SPECT in skull-base invasion in nasopharyngeal carcinoma is equivalent to that of MRI. The consistency between SPECT and MRI is good. Moreover, there is a good correlation between SPECT and symptoms, signs and stage. SPECT of skullbase tomography is necessary for patients with severe headache, negative CT and those who can not receive MRI. When SPECT result is positive,skull-base should be considered to be invaded and should be defined as gross tumor volume in radiotherapy planning.

Objective To promote the rational antibacterial drug use for common digestive system (DS) diseases by reviewing the clinical application of antibacterial drugs for hospitalized patients, formulating the hospital clinical specification of antibacterial drugs for common DS diseases and establishing the clinical application of antibacterial drug differentiation index for DS department.Methods 300 cases were selected from 2 060 hospitalized DS patients in the year 2014 based on the given disease ratio.The usage rate and intensity of antibiotics were calculated with survey and medical statistical analysis.The differentiation index for the clinical application of antibacterial drugs were established.Results Theoretical values of usage rate and use intensity of antibiotics in DS department were 29.0%, 41.5 DDDs according to analysis.However, the practical values were 40.3%, 50.3 DDDs.Conclusion It is necessary to improve the level of clinical antibacterial drug use.One effective way to achieve such goal is to establish the differentiation index of antibacterial drug clinical application for the department.

Objective To evaluate the protective effect of hydrogen-rich saline on renal ischemia/reperfusion (I/R) in mice. Methods Thirty C57BL/6 mice were randomly divided into 3 groups: sham-operated (SO) group, I/R group (mice were injected with 5 ml/kg saline by tail vein just before ischemia induction) and hydrogen-rich saline group (mice were injected with 5 ml/kg hydrogen-rich saline). At the 6th h after reperfusion, the sera and renal samples subject to IR injury were collected. The Scr and BUN levels in serum were determined and renal histological changes were also examined. The apoptosis of renal tubular epithelial cells was measured by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay. Malondialdehyde (MDA) contents in renal samples were measured using specific kits. The infiltration of F4/80 positive macrophages and neutrophils was assayed by using immunohistochemistry. The mRNA expression of TNF-α, IL-6, IL-1β and IL-17 was detected by using real time reverse transcription PCR. Results As compared with LR group, at the 6th h following reperfusion the levels of Scr and BUN were significantly reduced (P<0.05), histological changes obviously alleviated (P<0.01), apoptosis of renal tubular epithelial cells and MDA contents was decreased (P<0.05) in hydrogen-rich saline group. Moreover, the infiltration of macrophages and neutrophils, and the mRNA expression of TNF-α, IL-6, IL-1β and IL-17 in renal tissue in hydrogen-rich saline group were also declined as compared with IR group (P<0.05). Conclusion Hydrogen-rich saline can ameliorate renal IR injury to some extent, which is associated with inhibition of inflammatory response induced by reperfusion.