Background Diabetic retinopathy (DR) is one of the most important microvascular complications of diabetes,which has become one of the leading causes of blindness.Neovascularization is the main pathological manifestations of DR,but its mechanism is unknown.There is a clear need to investigate its pathogenesis which can offer potential therapeutic targets.Objective The aim of this study was to investigate the expression and distribution of visfatin and vascular endothelial growth factor (VEGF) in diabetic model rats.Methods This study was approved by Animal Ethic Committee of Inner Mongolia Medical University.Sixty SPF 8-week-old male SpragueDawley rats were randomized into the diabetic group and control group.The rats were housed under a condition that alternated between 12 hours of light and darkness,with free access to rat food and water.Diabetes was induced by intraperitoneal injection of 60 mg/kg (0.60 ml/100 g) of streptozotocin (STZ) and control rats received equivalent volume of buffer.The models were regarded as successful when blood glucose was ≥ 16.7 mmol/L.Rats were sacrificed 12 weeks after the injection of STZ and retinal specimens were prepared to detect the expression of visfatin and VEGF.Total retinal protein was isolated from the retinas of experimental and control eyes,and the expression of visfatin and VEGF was assessed by Western blot.Frozen cross sections of retinas of 5 μm thickness were used to perform double immunofluorescence staining with anti-visfatin and anti-VEGF antibodies.Results Mean body weight of the diabetic rats was (189.02±11.34) g and that of the control rats was (489.57 ± 14.48) g at 12 weeks post-injection,showing a significant difference between them (t =5.236,P =0.003).Mean blood glucose level was (29.25±3.86) mmol/L in the diabetic group and (5.32±1.01) mmol/L in the control group,demonstrating a significant difference (t =11.778,P =0.000).Double immunofluorescence staining showed reduced expression of visfatin and VEGF in the retinal nerve fibrous layer and glial cells in the control rats.A stronger staining for visfatin and VEGF was found in the various layers of the retina in the diabetic rats,with an expression level of visfatin (A value) of 346.26±41.23,which was considerably higher than that of the control group (102.07±65.01) (t =8.291,P =0.000) in 12 weeks after injection.Furthermore,the expression of VEGF in the retina was elevated in the diabetic group compared with the control group (A value) (415.88±92.15 vs.113.06±32.06) (t=10.067,P=0.000).Conclusions Visfatin might contribute to the pathologic progression of diabetic retinal,neovascularization and it might play a synergistic role with VEGF in the pathophysiology of DR.

Objective To study the influence of exposure factors on posttraumatic stress disorder( PTSD) and Posttraumatic Growth( PTG) in middle school students in disaster area four years after the Wenchuan earth?quake . Methods 1 526 students from four schools in Worst?Hit Areas were investigated with Self?compiled Earth?quake Exposure Factors Questionnaire,Posttraumatic Growth Inventory( C?PTGI) and Impact of Event Scale( IES?R). Data were analyzed by ANOVA and multiple linear regression analysis.Results The score of IES?R had sig?nificant difference between different levels of all exposure factors(F=5.75~89.10, P<0.05) ,and students with high exposure level((26.68±14.66),(26.80±15.56),(27.83±14.62),(29.02±15.36),(27.77±15.74),(26.74± 15.63),(25.43±14.32),(29.51±14.36)) had heavier symptoms of PTSD than those with low exposure level ((22.84±13.96),(23.98±13.99),(23.63±14.21),(23.53±13.96),(23.64±13.83),(24.24±14.15),(21.27± 14.35),(17.54±13.34)). Only exposure factors of having witnessed someone injured and having close friends se?riously injured or being killed could significantly influence the score of PTGI(F=11.82, P=0.001;F=6.23, P=0.013). Regression analysis showed that five exposure factors (grade,having felt scared,having family members being killed,having close friends seriously injured or being killed,having witnessed someone injured) had signifi?cant effect on IES(ΔR 2=0.141) ,but only one factor( having witnessed someone injured) had weak effect on PTG (ΔR 2=0.007).Conclusion Exposure factors can predict posttraumatic stress symptoms in middle school students in Wenchuan four years after the earthquake,and the emotion of fear is a strongest predictor,but they can not pre?dict posttraumatic growth.

Objective:To explore the predictive value of computed tomography (CT) texture analysis for the recurrence in children with acute pancreatitis (AP).Methods:The clinical diagnostic test was conducted. The clinical data of 56 children with primary AP who were admitted to Wuhan Fourth Hospital from January 2016 to January 2018 were collected. There were 13 males and 43 females, aged from 3.5 to 13.0 years, with a median age of 5.5 years. Based on follow-up in other hospitals, 20 children with recurrence of AP were allocated into recurrence group, and 36 children without recurrence were allocated into non-recurrence group. All the 56 children underwent abdomen plain and enhanced CT scan within 24 hours after first admission. Observation indicators: (1) comparison of clinicopathological features between two groups of children with AP. (2) comparison of CT texture parameters between two groups of children with AP. (3) diagnostic efficacy of clinical features and CT texture parameters. Follow-up using outpatient reexamination and telephone interview was conducted to detect recurrence of AP up to February 2020. The duration of follow-up required more than or equal to 24 months. The Shapiro Wilk test was used to analyze normality of measurement data. Measurement data with normal distribution were repre-sented as Mean± SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M (range) or M ( P25, P75), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was analyzed using the chi-square test. Comparison of ordinal data was conducted using the Mann-Whitney U test. Clinical parameters and CT texture parameters with statistical differences were multivariate analyzed using the Logistic regression model. Receiver operating characteristic curve (ROC) analysis was used to evaluate the predictive efficacy of parameters for recurrence of AP. Results:(1) Comparison of clinicopatholo-gical features between two groups of children with AP: cases with or without complications were 14 and 6 for the recurrence group, versus 7 and 29 for the non-recurrence group, showing a significant difference between the two groups ( χ2=14.021, P<0.05). Cases with minimal, moderately severe or severe disease (severity of disease) were 2, 5, 13 for the recurrence group, versus 19, 11, 6 for the non-recurrence group, showing a significant difference between the two groups ( Z=5.414, P<0.05). (2) Comparison of CT texture parameters between two groups of children with AP: the energy value in the arterial phase on CT examination was 0.186(0.174,0.206)for the recurrence group and 0.413(0.405,0.425) for the non-recurrence group, showing a significant difference between the two groups ( Z=9.413, P<0.05). The energy value and entropy value in the venous phase on CT examination were 0.084(0.078,0.092) and 0.961(0.210,1.720) for the recurrence group, versus 0.135(0.124,0.156) and 0.372(0.210,0.535) for the non-recurrence group, showing significant differences between the two groups ( Z=4.763, 7.243, P<0.05). (3) Diagnostic efficacy of clinical parameters and CT texture parameters: results of multivariate analysis showed the complications, severity of disease, energy value in the arterial phase on CT examination were related factors for recurrence in children with AP, energy value and entropy value in the venous phase on CT examination were related factor for recurrence in children with AP ( odds ratio=0.874, 0.765, 0.837, 0.902, 0.813, 95% confidence interval as 0.802?0.985, 0.581?0.914, 0.753?0.897, 0.862?0.948, 0.765?0.873, P<0.05). Results of ROC analysis showed that that areas under curve (AUC) of complications, severity of disease in the clinical parameters were 0.734 and 0.832, the AUC of single CT texture parameter was 0.811?0.867, the AUC of clinico-pathological parameters combined with CT texture parameters was 0.882. Conclusion:CT texture analysis can early and non-invasively predict the recurrence of AP in children, and the combination of clinicopathological parameters with CT texture parameter has a better predictive efficacy.

Objective To establish the optimal extracted method for content dete rmination of naringin in Citrus grandis. Methods RP-HPLC was used to determinat e the content of naringin extracted with the above two methods from different ye ar samples of Citrus grandis. Results The average content extracted with ultraso nic extraction was 13.53 %,and the average content extracted with soxhlet extr action was 11.98 %,there being insignificant difference between the two method s. Conclusion The content of naringin extracted with ultrasonic extraction is mo re than that with the soxhlet extraction,which be receipted in Chinese pharmeco pia. And ultrasonic extraction method is more convenient and can save time.

AIM: To analyze the effect of pigment epithelium derived factor (PEDF) on nitrogen monoxide (NO) and expression of cysteine-containing, aspartate-specific proteases-3 (caspase-3) in retinal tissues of model rats with optic nerve crush injury.METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group.Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples.After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF (0.2μg/μL).Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining.The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot.RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly.Retinal ganglion cells (RGCs) arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries.The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining.RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group.Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group (all P < 0.05).CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.

The focus of microbiologists has moved to the rare actinomycetes.For selective isolation of rare actinomycetes that all play the important role in bioactive compounds,the approaches which involve the methods using gellan gum and flooding solution、 rehydration-centrifugation(RC)、 extremely high frequency radiation(EHF)、 bacteriophage and sucrose-gradient centrifugation were introduced in this paper.

OBJECTIVETo observe the characteristics of sperm single-stranded DNA breaks (SSB) and double-stranded DNA breaks (DSB) in infertile men, explore the association of DSB with male infertility, and provide a new observation index and idea for the diagnosis and treatment of the disease.

METHODSThis study involved 60 infertile men (infertility group) and 30 normal healthy males with infertile wives (control group). We comparatively analyzed the seminal parameters of the two groups, determined sperm concentration and viability using the computer aided sperm analysis system, measured the sperm survival rate by hypoosmotic swelling (HOS) test, examined sperm morphology by Diff-Quick staining, and detected sperm DNA damage by two-tail comet assay.

RESULTSNine two-tail comet models were established for detecting sperm DNA integrity. Comparisons between the fertility and control groups showed that the sperm DNA fragmentation index (DFI) was (33.8 ± 13.1) vs (16.3 ± 7.9)% (P < 0.01), the SSB-DFI was (19.2 ± 11.4) vs (14.9 ± 7.6)% (P > 0.05), the SSB-DFI/DFI was (56.8 ± 32.4) vs (91.4 ± 27.8)% (P < 0.01), the DSB-DFI was (23.9 +13.4) vs (6.1 ± 2.7)% (P < 0.01), and the DSB-DFI/DFI was (70.8 ± 19.5) vs (37.4 ± 11.3)% (P < 0.01). The optimal cut-off value of DSB-DFI/DFI in the diagnosis of male infertility was 39.5%, with the AUG, sensitivity, and specificity of 0.969, 98.3%, and 90%; that of DSB-DFI was 15.85%, with the AUC, sensitivity, and specificity of 0.912, 86.7%, and 80%; and that of DFI was 18.65%; with the AUC, sensitivity, and specificity of 0.861, 90%, 70%, respectively. In the infertile men, neither SSB-DFI nor SSB-DFI/DFI exhibited any correlation with semen parameters (P > 0.05); DFI was correlated negatively with the percentage of progressively motile sperm, sperm survival rate, and the percentage of morphologically normal sperm (P < 0.05 or P < 0.01), but not correlated with sperm concentration (P > 0.05); both DSB-DFI and DSB-DFI/DFI showed a negative correlation with sperm concentration, sperm survival rate, and the percentages of progressively motile sperm and morphologically normal sperm (P < 0.05 or P < 0.01).

CONCLUSIONDouble-stranded, rather than single-stranded DNA breaks, may be a factor inducing male infertility. The type of sperm DNA strand damage is of much reference value for the assessment of male fertility.

Case-Control Studies ; Comet Assay ; DNA Breaks, Double-Stranded ; DNA Breaks, Single-Stranded ; DNA Fragmentation ; Fertility ; Humans ; Infertility, Male ; diagnosis ; genetics ; Male ; Semen Analysis ; Sensitivity and Specificity ; Sperm Count ; Spermatozoa ; Staining and Labeling

Background With the development of phacoemulsification surgery,the minimization of the size of the incision in order to reduce the damaging of eye tissue and postoperative complications becomes a focus.Objective This study aimed to assess the clinical efficacy of coaxial phacoemulsification type cataract surgery through a 1.8 mm microincision with foldable intraocular lens (IOL) implantation in eyes with age-related cataract.Methods Informed consent was obtained from each patient at the beginning of this study.A serial case observation study was designed.Thirty-two eyes of 32 patients with age-related cataract were enrolled in the Inner Mongolia Autonomous Region People' s Hospital from April 2012 to May 2012,including 8 eyes of grade Ⅱ hard nucleus cataract,16 eyes of grade Ⅲ hard nucleus cataract and 8 eyes of grade Ⅳ hard nucleus cataract.Ocular axial length was measured by an A-mode ultrasonic apparatus,and IOL diopter was calculated using the SRK-Ⅱ formula.Under ocular surface anesthesia,a 1.8 mm clear corneal tunnel incision was made at the 10-11 o' clock position,and then an auxiliary incision was made at the 2 o' clock position.The opaque lens was extracted by routine phacoemulsification.An Akreos MI60 IOL was implanted through the 1.8 mm incision.The time and level of ultrasonic power required for phacoemulsification,postoperative visual acuity and incidence of postoperative complication were assessed.Postoperative examinations were scheduled at 1 day,1 week and 1 month after the surgery.Results All the operations proceeded smoothly.The mean phaco-time was (7.0±3.6) seconds and the mean ultrasonic power level was (15.3 ±6.1)％ among the different grades of cataract groups.The number of eyes presenting an uncorrected distance visual acuity of ≥ 0.5 was 18 (56.25％),28 (87.50％) and 30 (93.75％) 1 day,1 week and 1 month after surgery,respectively.The number of eyes with a best corrected distance visual acuity of ≥ 0.5 and 0.8 were 31(96.88％) and 26 (81.25％),respectively,1 month after surgery.Negligible changes were detected in the anterior chamber depths,and no thermal damage was found at the incision during the operation.Conclusions Coaxial phacoemulsification and IOL implantation through a 1.8 mm microincision is safe and effective.Thorough planning and precise execution are necessary.

Objective To establish high resolution, reproducible 2-dimensional electrophoresis (2-DE) profiles of invasive and non-invasive pituitary adenoma tissues and to identify differentially ex-pressed proteins between the invasive and non-invasive tissues. Methods The proteome from invasive and non-invasive pituitary adenomas tissues was dissected and analyzed by: (1) immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis, (2) silver staining, (3) imageMaster 2-D software analysis, (4) peptide mass fingerprint based (PMS) on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and (5) database comparison. Results High-resolution 2-D patterns of invasive and non-invasive pituitary adenoma tissues were successfully produced and re-peated 3 times for each sample. An average of 1080±24 and 1035±28 spots were detected for invasive and non-invasive pituitary adenoma tissues, respectively. Additionally, 975±45 and 918±56 spots were found to have an average matching rate of 90.3% and 88.7% for invasive and non-invasive tissues, re-spectively. The spot positional deviation was (1.563±0.259) mm for IEF and (1.088±0.206) mm for SDS-PAGE. A total of 99 spots of differential expression were matched between the invasive and non-in-vasive pituitary adenoma tissues. Thirty differential proteins, some of which were involved in the regula-tion of cells cycle and signal transduction, were initially characterized by PMS. Conclusion The acquisi-tion of well-resolved and reproducible 2-D patterns of invasive and non-invasive pituitary adenoma tissues and the identification of differentially expressed proteins provides a proteome database for invasive pituita-ry adenomas.