Background The pathogenesis of dry eye is complicated,hormone level is thought to be one of impact factors in the development of dry eye.The regulation of the synthesis process of metalloproteinases(MMPs) in tissue has been reported.However,the effects of hormone on expression of MMP-2 in lachrymal gland is not clear.Objective This study was to investigate the effects of estrogen and androgen on the expression of MMP-2 in lachrymal gland in ovariectomized rats,and explore the role of MMP-2 in dry eye.Methods Sixty-four 3-monthold clean female Wistar rats were randomized into control group(8 rats),sham operation group(8 rats)and experiment group(48 rats).Ovariectomy(OVX) was performed on the rats of experiment group,and only fat tissue of abdominal cavity was cut off in the rats of the sham operation group.After 5 months of OVX,the experimental rats were subdivided into model control group,vehicle group,estrogen and androgen systemic or topical utilization groups and 8 rats for each group.Six weeks after administration of the drugs,the lachrymal gland was obtained.The expression of MMP-2 mRNA in the lachrymal gland was detected by reverse transcription PCR(RT-PCR),β-actin mRNA was used as an internal control,and the expression of MMP-2 protein was detected by Western blot,GAPDH was used as protein loading control.The use and care of the rats complied with the ARVO Statement.Results The expression of MMP-2 mRNA was strongest in the systemic estrogen group and was weakest in the systemic androgen utilization group.A significant difference in the MMP-2 mRNA expression was found among the 8 groups(F=18.60,P＜0.01),and the MMP-2 mRNA was significantly higher in the model group than that of the normal control group(0.66±0.10vs.0.47±0.10)(q=3.01,P＜0.05).In addition,the MMP-2 mRNA was significantly higher in the systemic estrogen group compared with the model group (0.83 ±0.10 vs.0.66-0.10) (q =2.79,P＜0.05) ; while the expression of MMP-2 mRNA was significantly declined in the systemic androgen group in comparison with the model group(0.12±0.04 vs.0.66±0.10)(q=11.41,P＜0.01).The MMP-2 protein presented with a strongest expression in the systemic estrogen utilization group and a weakest expression in the systemic androgen utilization groups.The expression level of the MMP-2 protein in the lachrymal gland was significantly different among the 8 groups(F =7.28,P＜0.01).The MMP-2 in the model group was significantly higher than that of the normal group(0.55±0.13 vs.0.38±0.08) (q =2.39,P＜0.05),and that in the systemic estrogen group was increased in comparison with the model group(0.69±0.12 vs.0.55±0.13) (q =1.85,P＜0.05).However,the MMP-2 in the systemic androgen group was significantly lowed in comparison with the model group(0.27±0.07 vs.0.55±0.13) (q =4.32,P＜0.01).Conclusions Estrogen may up-regulate the expression of MMP-2 in lachrymal gland,but the effect of androgen is opposite.Hormone level may play an important role in the regulation of the function of lachrymal gland.

Actins ; metabolism ; Adenoma, Pleomorphic ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Infant ; Keratins ; metabolism ; Male ; Nasal Cavity ; pathology ; Neoplasm Recurrence, Local ; Nose Neoplasms ; metabolism ; pathology ; surgery ; Reoperation ; Vimentin ; metabolism

To explore the regulation of eIF4E, we screened the protein interacting with eIF4E from human cDNA library by using yeast two-hybrid system. Several clones interacting with eIF4E were identified. One of them was homologous with HUWE1 (HECT, UBA and WWE domain containing 1, also named as ARF-BP1, HECTH9 or HUWE1). Cell co-immunoprecipitation showed that eIF4E could bind to HUWE1 in mammalian cells. We also found that HUWE1 bearing the HECT domain is necessary for its association with eIF4E.
Animals ; Eukaryotic Initiation Factor-4E ; metabolism ; Humans ; Ubiquitin-Protein Ligases ; metabolism

AIM: To establish a HPLC method for determination of curcumol and germacrone in Zedoary Turmeric Oil and Glucose Injections. METHODS: RP-HPLC was applied for quantitative analysis of curcumol and germacrone. Hypersil ODS2 column (250 mm? 4.6 mm, 5 ?m) and gradient elution was used. Mobile phase A was acetonitrile and mobile phase B was water. Phase A time-percentage composition was as follows:0-3 min,45%;3-30 min,45%→65%;30-38 min, 65%; 38-45 min, 65%→90%;45-55 min,90%→100%;55-65 min,100%. The flow rate was at 1.0 mL?min -1 . Diode array detector was set at 214 nm and column temperature was at 25 ?C. RESULTS: The calibration curves of curcumol and germacrone were linear in the range of 0.26 - 6.5 ?g(r= 0.999 9 ) and 0.112 5 - 2.812 5 ?g(r=1), The average recoveries of them were 102.1% and 98.8% (n=5), respectively. CONCLUSION: Curcumol and germacrone can be determined by the method, and this will improve the quality control of Zedoary Turmeric Oil and Glucose Injections.

Objective To study antibiotic resistance phenotypes and genotypes of extended spectrum ?-lactamases (ESBLs) and AmpC ?-lactamase-producing Klebsiella oxytoca isolated from specimens of respiratory tract in children.Methods Bacterial isolates were identified by API or VITEK32. Agar dilution was used for antibiotic susceptibility test,and ESBLs and AmpC were detected by confirmatory test recommended by CLSI/NCCLS and by 3-aminophenylboronic acid (APB) disk potentiation test, respectively.Microarray was used to determine the genotypes of ESBLs and AmpC ?-lactamases.Genotypes of Klebsiella oxytoca were determined by enterobacterial repetitive intergenic consensus (ERIC)- PCR.Results ESBLs were positive in 129 out of 165 isolates (78.2%).Both ESBLs and AmpC ?- lactamases were positive in 16 out of 165 isolates (9.7%).AmpC ?-lactamase alone producer was not detected in term of phenotype and genotype.CTX-M was the most common type of ESBLs and DHA was the only type of AmpC ?-lactamase in these isolates.Most antibiotic resistant strains of Klebsiella oxytoca possessed the same genotype by ERIC-PCR.Although all strains were susceptible to carbpenem,Klebsiella oxytoca with ?-lactamases were more resistant to other antibiotic agents than those without ?- lactamases.Conclusions There is high prevalence of ESBLs production among Klebsiella oxytoca isolated from children in Urumqi.The main genotypes of ESBLs and AmpC ?-lactamases are CTX-M and DHA.

Norovirus ; chemistry ; Receptors, Virus ; physiology ; Viral Structural Proteins ; chemistry ; immunology ; Viral Vaccines ; immunology

China ; Congresses as Topic ; Fatty Liver ; Humans

In central nervous system only a limited number of vesicles exist in the presynaptic terminals. The size and fusion modes of the vesicles were particularly important because of their potential impact on neuronal communications. Efficient methods were needed to analyze the recycling kinetics of synaptic vesicle and the size of readily releasable pool (RRP). In this study, fluorescent dyes with different affinity for membranes (FM1-43 with high affinity and FM2-10 with low affinity) were used to stain the functional synaptic vesicles of cultured hippocampal neurons and the kinetics of vesicle recycling was measured. The results showed that the destaining proportion was larger for FM2-10 than that for FM1-43 during the first trial, while it was greater for FM1-43 than FM2-10 during the second and third trials (first round, 93.0%+/-5.9% versus 57.9%+/-3.5% for FM2-10 and FM1-43, respectively, P<0.0001; second round, 1.4%+/-3.8% versus 24.0%+/-2.3%, P<0.0001; third round, 2.3%+/-1.6% versus 8.6%+/-1.5%, P=0.005). The results indicated that rapid endocytosis existed not only in the first round but also occurred when the vesicles were reused. Moreover, Both high-frequency stimuli and hypertonic sucrose stimuli were used to estimate the RRP sizes in the mix cultured hippocampal inhibitory neurons at 13-14 days in vitro (DIV). We found that the RRP size estimated by hypertonic sucrose stimuli [(200+/-23.0) pC] was much larger than that estimated by high-frequency stimuli [(51.1+/-10.5) pC]. One possible reason for the discrepancies in RRP estimates is that in mix cultured conditions, one neuron may receive inputs from several neurons and hypertonic sucrose stimuli will cause RRP of all those neurons release, while using dual patch recording, only the connection between two neurons was analyzed. Thus, to exclude out the impacts of inputs numbers on RRP sizes, it is more reasonable to use high-frequency stimuli to estimate the RRP size in mix cultured neurons.
Cells, Cultured ; Endocytosis ; Hippocampus ; cytology ; Neurons ; physiology ; Synaptic Vesicles ; physiology

Objective To clone and express Dengue virus nonstructural protein 4A (NS4A) gene and express in eukaryotic cells.Then,to isolate and purify and isolate cellular proteins interacted with NS4A.Methods With specific primers,NS4A gene fragment tagged with FLAG and HA (FLAG-NS4A-HA) was amplified by PCR and cloned into an expression vector,pSG5 vector.Recombinant plasmid was transfected into A549 cells by LipofectAMINETM2000.Transient expression of FLAG-NS4A-HA was detected by Western blot.The NS4A interacting proteins were isolated and purified by tandem affinity purification (TAP) system using HA and FLAG antibodies,and then assayed by silver stained SDS-PAGE.Results Dengue virus NS4A gene tagged with FLAG and HA was successfully constructed into pSG5 vector and expressed in A.549 cells.Silver stained SDS-PAGE showed that the expressed NS4A and two potential interacting proteins that interact with NS4A were isolated after TAP purification and SDS-PAGE.Conclusion Cellular proteins that potentially interacted with Dengue virus NS4A were successfully purified and isolated,which provided a basis for further research.