Furbenicillin is a broad-spectrum semisynthetic penicillin with strong antibacterial activity against Gram-negative bacteria. Furbenicillin sodium is determined by volumetric method in current criteria. However, the criteria does not contain an assay of related substances of furbenicillin sodium. In this study, we established a method for detection and analysis of furbenicillin sodium and its related substances by HPLC. The analysis was performed with a C18 column under a gradient elution, the detection wavelength was 225 nm, and the column temperature was 35 degrees C. The reliability and accuracy of established method was validated in this study. Pure samples of furbenicillin sodium and its related substances were prepared. The structures, biological activities, and chromatographic retention behaviors of furbenicillin sodium and its related substances were identified using NMR, CLSI agar dilution method, and HPLC. All results in the current study provide ample evidence that this method is able to determine the reasonable limits in the quality-control protocol for furbenicillin sodium.
Anti-Bacterial Agents ; chemistry ; Chromatography, High Pressure Liquid ; Magnetic Resonance Spectroscopy ; Penicillins ; Quality Control ; Reproducibility of Results

Background Graft rejection is a primary cause of corneal transplantation failure,especially in high-risk keratoplasty.How to extend the survival time of graft is a problem to be solved.Objective This study was to investigate the influence of immune tolerance on high-risk rat keratoplasty induced by donor bone marrowderived immature dendritic cells (imDCs) transfected by chemokine receptor 7 (CCR7) recombinant adenovirus (Ad).Methods Bone marrow-derived imDCs were isolated and cultured from femur marrow of one male Wistar donor rat.The cells were transfected using recombinant Ad vector with rat CCR7 gene and resuspended in 500 μl PBS containing 1％ fetal bovine serum with the cells 1 × 107.High-risk corneal transplantaion models were established using monolateral corneal alkali-burn method in 60 SD rat recipients, and then allograft keratoplasty was performed with the 30 Wistar rats as donors.The models were randomized into the PBS group,imDCs group,imDCs with blank Ad vector group and imDCs with Ad-CCR7 group following the corresponding solution injection via caudal vein on preoperative day 7 and postoperative day 3 respectively.The survival time of graft was evaluated under the slit lamp microscope once per day.On the 14th day after operation, corneal sections were prepared from 6 eyes of each group for the pathological examination,and the relative expression levels of T helper cell 1 (Th1)-related factors,interleukin-2 (IL-2) mRNA and interferon-γ(IFN-γ) mRNA,as well as Th2-related factors, IL-4 mRNA and IL-10 mRNA, were detected by reverse transcription PCR (RT-PCR).The use and care of animals complied with the ARVO statement Results The mean survival time of grafts was (10.44±1.88) , (16.00±2.18) , (15.11±2.03) and(23.67±2.83) days in the PBS group,imDCs group,imDCs with blank Ad group and imDCs with Ad-CCR7 group, respectively, with a significant difference among the 4 groups (F =53.005, P =0.000).Compared with the PBS group, the mean survival time of grafts was considerably extended in the imDCs group,imDCs with blank Ad group and imDCs with Ad-CCR7 group (t=5.220,4.385,12.423 ,all at P=0.000) ,and a remarkble prolongation of graft survival duration was seen in the imDCs with Ad-CCR7 group in comparison with the imDCs group and the imDCs with blank Ad group (t =7.204,8.039,both at P=0.000).The relative expression levels of IFN-γ mRNA and IL-2 mRNA in the grafts were significantly lower,but the relative expression levels of IL-4 mRNA and IL-10 mRNA were significantly higher in the imDCs group,imDCs with blank Ad group and the imDCs with Ad-CCR7 group than those in the PBS group (all at P＜0.05).Compared with the imDCs group and the imDCs with blank Ad group, the relative expression levels of IFN-γ mRNA and IL-2 mRNA in the grafts were remarkably delined,but the relative expression levels of IL-4 mRNA and IL-10 mRNA were remarkably elevated in the imDCs with Ad-CCR7 group (all at P =0.000).Conclusions Application of imDCs transfected with CCR7 recombinant Ad via vena caudalis can prolong the survival time of grafts after keratoplasty of SD rats probably by affecting the balance of Th1/Th2 cytokines.

AIM: To evaluate the safety, efficacy and stability of posterior chamber phakic intraocular lens ( ICL ) implanation and clear lens extraction for the correction of high myopia.
METHODS: The study enrolled 56 cases ( 100 eyes ) of high myopia. Group I comprised 32 cases ( 58 eyes ) receiving ICL implantation and Group II comprised 24 cases (42 eyes) undergoing clear lens extraction. In this study, we evaluated the two groups of subject's the visual and refractive results, intraocular pressure ( IOP ) , endothelial cell density ( ECD ) , anterior chamber depth ( ACD) , lens transparency, the surgical complications as well as visual adverse symptoms before and after surgery.
RESULTS: The postoperative subjects in group I and group II were followed, uncorrected vision acuity ( UCVA)>0. 5 were 69. 0% in group I and 71. 4% in group II after 3mo. UCVA>0. 5 were 72. 4% in group I and 73. 8% in group II after 1a. Predictability of the manifest spherical equivalent refraction within±1. 00D was achieved in 62. 1%of eyes in group I and 57. 1% in group II after 1a. The central vault of the ICL ( distance from posterior surface of ICL to the crystalline lens ) measured with anterior segment optical coherence tomography ( AS-OCT ) was 0. 35-0. 54 (0. 40±0. 16) mm. Twelve point one percent of eyes in group I and 7. 1% of eyes in group II had transient mild increase in IOP. Here were statistically significant differences between preoperative and postoperative ECD (P<0. 001 ). Complications of surgery: 1 eye had ICL spontaneous rotation, 2 eyes had anterior subcapsular cataract, 4 eyes noticed halos around lights at night in group I. Three eyes had posterior capsule mild opacification, 3 eyes noticed halos around lights at night, 12 eyes had difficulty in near vision in group II.
CONCLUSION: ICL implantation and clear lens extraction are effective, safe and predictable surgical option for the management of high myopia. No severe complications occurred, but its long time effect and safety still need more time to prove.

Objective To study the composition of variant and point mutations of Norovirus GⅡ.4 in Guiyang regions.Methods From June to November 2010,cases information and fecal specimens were collected from guard-hospitals in Guiyang regions,who had caught the acute-gastroenteritis.Noroviruses in specimens were detected by a real-time reverse transcription polymerase chain reaction(real-time RT-PCR),and then partial genotyped norovirus-positive clinical samples (in random) were cloned and sequenced in VP1 gene code.Furthermore,the gene sequences were compared with the published variants at home and abroad of norovirus(GⅡ.4),including the phylogenetic analyses of genomes and variation of amino acids within individual sites.Results Those 267 specimens were GⅡ-norovirus-positive(62.68％) in 426 clinical samples.There were nine GⅡ.4-norovirus-positive VP1 gene-sequences available,and two subtype-norovirus variants (GⅡ.4 2008a and G Ⅱ.4 2008b variant) were epidemic in 2010,Guizhou province.The homology between and in subgroups were 95.90％-96.72％ and 99.45％-100％.Two amino acids within individual sites were apt to mutate.Conclusion Norovirus GⅡ genotype were predominant in summer and fall acute gastroenteritis in 2010 for Guiyang regions,and the variants were diversity.

Objective To construct recombinant adenovirus vector containing mouse CD40Ig fusion gene for the study of induction of donor-specific tolerance. Methods CD40Ig fusion gene was constructed by PCR overlapping technique, and was cloned into the shuttle plasmid pAdTtrack-CMV. The linearized shuttle plasmid was co-transfected into the E.coli strain BJ5183 with bone plasmid pAdeasy1, then the recombinant adenovirus plasmid was generated. The adenovirus was packaged and amplified in Cells 293. Results The recombinant virus AdCD40Ig was successfully constructed and its titer was 5?109 efu/ml. Conclusion AdCD40Ig can be used as an agent in experiment to induce donor-specific tolerance.

Objective To explore the structure of violent attitude and the predictive character of the passionate criminal and recidivist. Methods This study used FMMU-Abnormal Personality Risk Factors Inventory (FMMU-APRI) for measuring explicit violent attitude of the passionate criminal and recidivist in prison; while used the picture Attitude Activation paradigm (AAP) for assessing implicit violent attitude. Results The AAP results showed that parts of passionate criminal and recidivist had reversed priming effect, according to whether reversed, there were divided into four types: no-reversal passionate criminals, reversal passionate criminals, no-reversal recidivists and reversal recidivists, VIO scores of no-reversal passionate criminals and recidivists recidivist were significantly higher than the norm, as reversal passionate criminals and recidivists had no significance; in noreversal passionate criminals, compatible response rate was 1.104, incompatible response rate was 1.053; in reversal passionate criminals, compatible response rate was 1.042, incompatible response rate was 0.997; in no-reversal recidivists, compatible response rate was 1.059, incompatible response rate was 1.097; in reversal recidivists , compatible response rate was 1.039, incompatible response rate was 0.998, as each group had strong priming effect, and each group had a dissociation of implicit and explicit violent attitude. Conclusion The results suggest that offenders who have committed a crime in the same category also had a different violent attitude; and integrating indirect methods with direct methods would predict a crime more accurately.

Objective To evaluate the influence of two different education approaches on diabetic knowledge of type Ⅱ diabetes.Methods 79 type Ⅱ diabetes cases were collected from March 2010 to May 2010,and divided into experimental group and control group according to admission time.Experimental group was those in the first-half,and received figure dialogue education.Control group was those in the last-half,and received traditional heath education.Diabetes knowledge assessment questionnaire was used to evaluate knowledge level before and after education.Results Knowledge level was higher after education than that before education in both groups.No differences was found between the two groups ( P ＞ 0.05 ).All subjects in experimental group thought figure education could improve the study interesting,68.3％ subjects thought mastering knowledge about diabetes was useful,and 80.0％ subjects expect discussion by group.However,74.4％ subjects in control group thought traditional education could improve the study interesting,15.4％ subjects thought mastering knowledge about diabetes was useful,and 46.2％ subjects expect discussion by group.Conclusions Compare with traditional education,figure education although could not increase the knowledge level about diabetes significantly,but could highly increase the interesting of study.

Hydrolytic amino acids were extracted by acid hydrolysis method, then derivatized with phenyl isothiocyanate (PITC). And the samples were analysed by HPLC on an Ultimate Prime C18 (4.6 mm x 250 mm, 5 microm) column with gradient elution of 0.1 mol x L(-1) sodium acetate buffer solution (adjusted to pH 6. 5)-acetonitrile (93:7) (A) and acetonitrile-water (8:2) (B) at a flow rate of 1.0 mL x min(-1). Column temperature was 40 degrees C and the detected wavelength was 254 nm. Amino acids derivative solution remained stable in 36 hours. The response was linear for 16 amino acids with a correlation coefficient r > 0.999 5. The average recoveries were 98.01% -101.8%. The method is reliable with good accuracy and repeatability, which is useful for the determination of amino acids in Galli Gigerii Endothelium Corneum.
Amino Acids ; analysis ; Animals ; Chickens ; Chromatography, High Pressure Liquid ; Chromatography, Reverse-Phase ; Endothelium ; chemistry ; Gizzard, Avian ; chemistry

Objective To observe the changes of bone mass in reloaded rats after tail-suspension,and the effect and mechanism of simvastatin on this process.Methods Twenty-four 5-month old rats were divided into 4 groups of 6 animals in each group: Control (CL) group without tail-suspension,unloaded (UL) group with tail-suspension for 6 weeks,other 12 rats received tail-suspension for 3 weeks,then reloaded for subsequent 3 weeks (UL+RL) or combined with simvastatin treatment (UL+RL+SIM) at a dose of 10 mg/kg/d.All rats were sacrificed 6 weeks later,and the left femur was used for examination of bone mineral density,left tibia was used for bone histomorphometry analysis,the right femur and tibia were harvested for biomechanical test,and expression levels of type I collagen by real-time PCR and Western blot,respectively.Results 1.BMD of the CL group was significantly higher than those of the other three groups (P<0.05),and was markedly lower than those in the UL+RL and UL+RL+SIM groups (P<0.05).2.The bone histomorphometry showed that BV/TV in the CL group was significantly higher than those in the other 3 groups,and the UL+RL and UL+RL+SIM groups showed a significantly higher BV/TV than that of UL group (P<0.05).The Tb.Th was significantly higher in the CL group than in the UL group.The Tb.Sp in the CL group was significantly lower than those in the other 3 groups (P<0.05).The UL+RL and UL+RL+SIM groups showed significantly lower Tb.Sp than that of the UL group (P<0.05).3.Biomechanical test showed that the maximal load and elastic modulus in the CL groups were significantly higher than those of the other three groups (P<0.05).4.Real-time PCR showed that no significant difference in the mRNA expression level of Col I was found between any two groups.5.Western blot showed that the IOD of Col I is significantly lower than that in the CL group.Conslusions Bone loss,destruction of trabecular bone micro-architecture and biomechanical properties and reduction of type 1 collagen are present in tail-suspension treated rats,which are partially restored after reloading,and this recovery process is not enhanced by simvastatin treatment.

A hybridoma cell line(B10)was established by fusing spleen cells of BALB/c mou- se immunized with human thyroglobulin(HTG) with SP2/0 myeloma cells. An averagefusing rate of 6.3% and antibody-secreting positive well rate of 58.3% were obtained.During the first two months, the supernatant of B10 culture had a titer of 1/128 to1/2048 measured by hemagglutination method, and the ascites was positive at 1/64000-128000 and 1/320000 respectively as measured by hemagglutination and radioimmunoass-ay.The B10 cell line is very stable and has very high activity to produce anti-HTGmonoclonal antibodies. After several times of preservation in liquid nitrogen andpassage in culture for one year,a recent determination shows that cell culture super-natant and ascites still have very high titer,being 1/4096 and 1/1048576 respectively asmeasured by hemagglutination method. The chromosome number of the B10 hybridomacell is 99.5?7.4.The success of the establishment of this cell line is briefly discussed.Attempt to establish diagnostic kit with this monoclonal antibody is being undertaken.