Background Studies indicate that the decline of androgen level affects the secretion function of epithelial cells of meibomian glands and lacrimal glands ,which results in the abnormality of tear quality and quantity.However,whether androgen level regulates the mucin adhesion to ocular surface is still not elucidated.Objective This study was to investigate the tear functional changes and corneal ultrastructural changes in castrated male mice.Methods Fifty-six SPF male BALB/c mice were devided into normal control group, sham group, and orchectomy 1-week group, orchectomy 2-week group, orchectomy 4-week group, orchectomy 6-week group, orchectomy 8-week group according to random number table.Orchectomy was performed in the mice of the orchectomy 1-,2-,4-,6-and 8-week group,and only incision rather than enucleation of testis was carried out in the mice of the sham group.Peripheral venous blood samples were collected during eyeball enucleation in all the mice and serum androgen concentration was detected by radioimmunoassay.Tear secretion level was measured with phenol red thread.Break-up time of tear film (BUT) was tested,and corneal inflorescence staining was scored based on the criterion of Toshida.The corneas of all the mice were collected for ultrastructural examination under the transmission electron microscope.The study complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results Mouse serum androgen concentration was (573.87±6.74) ng/μl in the normal control group and (579.74-±8.24)ng/μl in the sham group, and there was no significant difference between these two groups (t =1.432, P =0.251) , however, the serum androgen concentration was 0 ng/μl in all castrated mice after orchectomy.The tear secretion levels were (5.26 ± 0.99), (4.89 ± 0.56), (4.62 ± 0.83), (4.28 ± 0.63), (3.36 ± 0.56),(2.60±0.27) and (2.08 ±0.35) mrn/5 minutes in the normal control group, sham group and orchectomy 1-, 2-, 4-,6-and 8-week groups, respectively, and the tear secretion levels were significantly lower in the orchectomy 2-, 4-,6-and 8-week groups than those in the normal control group (all at P＜0.05).The BUT were (68.33±12.86), (62.47± 3.75),(58.67±10.03), (47.17±7.64) ,(39.51±3.39),(32.67±3.88) and (31.00±2.36) seconds in the normal control group, sham group and orchectomy 1-, 2-, 4-, 6-and 8-week groups, respectively, and the BUTs were significantly shorter in the orchectomy 2-,4-, 6-and 8-week groups than those in the normal control group (all at P＜0.05).The fluorescein stain score of corneal epithelium was gradually increased with the lapse of castrated time.The decrease of number,shortening and edema of microvilli as well as separation of desmosomes of corneal epithelial cells were found under the transmission electron microscope in castrated mice.Conclusions Decreasing of serum androgen level, declining of aqueous tears,instability of tear film and damage of corneal epithelial cell ultrastructures occur in castrated male mice, and these manifestations are similar to dry eyes.

Objective:To measure the safe range from the foramen rotundum to the siphon of internal carotid artery using three-dimensional reconstruction technique, and to provide guidance for operation through pterygopalatine fossa.Methods:The skulls of 121 volunteers were scanned to get the final results with thin-section computed tomographic images.The position of the siphon of internal carotid artery (point A)and foramen rotundum (pointB)were ascertained.Three-dimensional reconstruction technique was used to build a coordinate system paralleled to the frankfort horizontal plane and the nasal septum plane.The coordinate system took point A as the coordinate origin.Point C and point D were the projections of point A in two planes parallel with frankfort horizontal plane and nasal septum plane which included point B.The distances of AC,AB,and BC were measured. The angles of the line went through A and B to the three planes were also measured.Results:The distance of AC was measured as 13.22 (3.79)mm (range,8.33 - 105.67 mm;95%CI:8.55 - 21.39 mm).The angle to the sagittal plane was measured as 33.54 (9.23)° (range,5.38- 66.58°;95%CI:30.88 - 34.20°). The angle to the coronal plane was measured as 53.17 (10.48)°(range,5.60-75.02°;95%CI:51.29-55.06°).The angle to the horizontal plane was measured as 9.43 (12.91 ) mm (range,- 28.44 - 82.22;95% CI:7.11 - 11.76 ). Conclusion:The safety distance from foramen rotundum to the siphon of internal carotid artery in the operation through pterygopalatine fossa (PPF) under nasoendoscope is obtained by thin-section computed tomographic images.

Objective To explore the serum anti-Mullerian hormone (AMH) level in women of childbearing age with normal menstrual cycles. Methods A total of 1 423 women with regular menstrual cycles were selected and divided into 5 groups according to their ages, i.e.≤25, 26-30, 31-35, 36-40,≥41 years. Their serum levels of AMH were measured, and the relationship between AMH and age was analyzed. Results The serum AMH levels of 5 groups according to ages (≤25, 26-30, 31-35, 36-40, ≥41 years) were 3.62, 3.10, 2.27, 1.07, 0.45μg/L, respectively. The comparison of serum AMH levels in different age groups had significant difference (P<0.01). Serum AMH level declined with increasing age,and dropped significantly after 36. The serum AMH level and age showed a negative correlation with significant difference (r=-0.374, P<0.01). Quadratic regression of logAMH proximally reflected the relationship between AMH and age. Conclusion AMH determination for women of childbearing age could provide reference for the evaluation of ovarian function.

Objective To research the jugular foramen,internal auditory pore (IAP)and the turning point between its components by imageological methods and to provide theoretical basis for retrosigmoid approach in the operation of acoustic neurinoma.Methods The skulls of 100 volunteers were scanned to get the final result with thin-section computed tomographic image. High-resolution spiral CT multiplane reformation was used to reform images that were parallel to the Frankfort horizontal plane to measure the distance between the turning point of retrosigmoid (A),the edge of jugular foramen(B)and the lower edge of the internal auditory canal(C)(denoted AC,AB,BC) and the shortest distance from the jugular foramen to AC.The angles between AC,AB and sagittal axis(α,β) were measured.Results The distance of AC was (44.94 ± 3.84)mm,the distance of AB was (43.68 ± 4.56)mm.The distance of BC was (6.15 ±2.04)mm,and the shortest distance between jugular foramen and AC was (5.21±0.23)mm.The angleαwas measured as (39.50±4.74)°,and the angleβwas measured as (46.35± 5.51)°.Conclusion The research measure the distance and angle between entry points and landmarks of retrosigmoid approach and the safe distance.

Objective:To explore the difference in ocular surface microbiota between patients with and without dry eye.Methods:Forty-two patients (42 eyes) diagnosed with dry eye were enrolled as dry eye group in the First Affiliated Hospital of Xi'an Jiaotong University from June to November 2020, and 37 controls without dry eye (37 eyes) were enrolled as control group in the same period.One eye was selected as the study eye, and the right eye was included when both eyes met the inclusion criteria.Swab samples from the conjunctival sac were obtained and sequenced.Sequencing of the V3-V4 region of the bacterial 16S rRNA gene was performed with Miseq PE301+ 8+ 301 platform.Operational taxonomic species (OTUs) clustering of microflora, comparison of alpha and beta diversity analysis of microflora between the two groups, annotation analysis of species and analysis of microbial markers were performed.The study protocol was approved by the Ethics Committee of The First Affiliated Hospital of Xi'an Jiaotong University (No.XJTU1AFCRC2018SJ-014). Written informed consent was obtained from each subject before any medical examination.Results:A total of 18 586 OTUs were obtained, and 3 674 OTUs were shared between the two groups.Alpha diversity analysis showed that there was no significant difference in observed species index, Chao index, Ace index, Shannon index and Simpson index between the two groups (all at P>0.05), suggesting there was no difference in microbiota richness between them.The PCoA analysis showed that the microbial compositions of the two groups were significantly different ( R2=0.039, F=3.100, P=0.022). The dominant flora of the two groups was similar, with Proteobacteria, Actinobacteria, Firmicutes, Bacteroidetes and Cyanobacteria as the top 5 abundant bacterial phyla, with Pelomonas, Corynebacterium, Propionibacterium, Pseudomonas and Herbaspirillum as the top 5 bacterial genera.LEfSe analysis identified Tissierellaceae, Enhydrobater and Finegoldia as dominant bacterial genera in dry eye group, and Caulobacter and Curvibacter in control group. Conclusions:The composition of ocular surface microbiomes is different between dry eye patients and controls.

OBJECTIVE:To screen the formulation of 5-fluorouracil (5-FU) polylactic acid (PLA) sustained-release discs (5-FU-PLA-DS),and study its in vitro drug-release mechanism. METHODS:UV spectrophotometry was used to determine the 5-FU content in the release medium. Using simulate body fluid as release medium,in vitro drug-release test was conducted under 37℃water bath. Using PLA with molecular weight of 3000,6000,10000,15000,20000,15 species of round 5-FU-PLA-DS with drug containing of 1.5,2.5,3.0 mg/piece and 3.0 mm in diameter and 1.0 mm in thickness were prepared. Using effective con-centration sustained release time and cumulative release rate as indexes, the optimal formulation was screened. The form of 5-FU-PLA-DS was observed by scanning electron microscopy after release,and its release mechanism was evaluated. RESULTS:In the optimal formulation, the PLA molecular weight was 20000 and drug containing was 3.0 mg/piece. The prepared 5-FU-PLA-DS can release for 119 d,with cumulative release degree of 100.63% and effective concentration sustained for 91 d. Scanning electron microscopy showed that the microspheres at the surface were degraded to the release medium first,and then the microspheres of inner layer exposed and release drug gradually after PLA degraded. The main mechanism of drug-release was melt-ing and diffusion. CONCLUSIONS:5-FU-PLA-DS is successfully prepared,with long release time in effective concentration,can be degraded step by step from outside to inside and achieve non-synchronous drug-release of microspheres at different layers.

In order to characterize a thermostable urate oxidase (Uox) from Microbacterium sp. strain ZZJ4-1, we cloned its gene (uox). The open reading frame of uox contained 894 base pairs and encoded a protein with 297 amino acids. Alignment of gene sequences indicated there was no obvious identity with the most reported uox and that 72% identity was found with uox from Arthrobacter globiformis. We inserted the gene into the plasmid pET-15b to construct an expression vector pET-15b-uox and got it induced expression in Escherichia coli BL21 (DE3). After the purification of the recombinant Uox by the HisBind column, we studied some properties of it. It was composed of subunits with a molecular mass of about 35 kDa. The optimal temperature and pH was 30 degrees C and pH 7.5. It was stable below 65 degrees C and from pH 8.5 to 11.0. The Km value was 0.22 mmol/L with the uric acid as the substrate. Ag+, Zn2+, CU2+ and SDS could totally inhibit its activity while Tween 20, Tween 80 and Triton X-100 had a slight promotion effect. The thermal stability of this enzyme was the most excellent among the reported recombinant Uox. Based on this property, it would be very useful in the application.
Actinomycetales ; enzymology ; genetics ; Amino Acid Sequence ; Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; Urate Oxidase ; genetics ; metabolism

Objective: To observe the inhibitory effects of nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) on vascular endothelial growth factor (VEGF) expression in hypoxia-induced human retinal pigment epithelial cells. Methods: The cultured APRE-19 cells were divided into Avastin group and VAS2870 group, and then the Avastin group was subdivided into the normoxic control group, hypoxia control group, 0.25 mg/ml Avastin intervention group, 0.50 mg/ml Avastin intervention group and 0.75 mg/ml Avastin intervention group, and the VAS2870 group was subdivided into 1 μmol/ml VAS2870 intervention group, 3 μmol/ml VAS2870 intervention group and 5 μmol/ml VAS2870 intervention group.CoCl₂ of final concentration of 300 mol/L was added to the medium to establish the cytochemical hypoxia model.The expressions of NOX4 and VEGF in human retinal pigment epithelial cells were located and evaluated by immunofluorescence staining, and relative expressing levels of NOX4 and VEGF proteins were compared by Western blot assay. Results: The relative expression of NOX4 was 0.657±0.153, 1.000±0.200, 1.206±0.300, 1.260±0.200 and 1.413±0.273, and the relative expression of VEGF-A was 0.821±0.110, 1.210±0.100, 0.672±0.100, 0.340±0.120 and 0.300±0.130 in the normoxic control group, hypoxia control group, 0.25 mg/ml Avastin intervention group, 0.50 mg/ml Avastin intervention group and 0.75 mg/ml Avastin intervention group, respectively, with statistically significant differences among the groups (F=17.631, P<0.001; F=4.777, P<0.05). The relative expression of NOX4 protein in 0.75 mg/ml Avastin intervention group was significantly lower than that in normoxia control group (P<0.001). The relative expression of VEGF-A protein in the cells of the 0.25, 0.50 and 0.75 mg/ml Avastin intervention group was significantly lower than that in hypoxia control group (P<0.05). The expression of NOX4 protein in the cells was 0.970±0.120, 1.060±0.130, 0.880±0.130, 0.567±0.135 and 0.450±0.120, and the relative expression of VEGF-A protein was 0.387±0.135, 0.627±0.125, 0.370±0.140, 0.363±0.140 and 0.160±0.100 in the normoxia control group, hypoxia control group, 1 μmol/ml VAS2870 intervention group, 3 μmol/ml VAS2870 intervention group and 5 μmol/ml VAS2870 intervention group, respectively, with statistically significant differences among them (F=12.933, P<0.001; F=4.948, P<0.05). The relative expression of VEGF-A protein in the 1, 3 and 5 μmol/ml VAS2870 intervention group was significantly lower than that in hypoxia control group (P<0.05). Conclusions NOX4 inhibitor can inhibit the expression of VEGF-A protein in hypoxia-induced human RPE cells by down-regulating the NOX4 level.