Background Lewis rat is a commonly used specie in experimental autoimmune uveitis(EAU).However,the characteristics of EAU,especially ocular uhrastructural change,are rarely reported.Objective This study was to investigate the inflammatory characteristics and ocular uhrastructure of EAU models in Lewis rat.Methods EAU models were induced in 12 SPF female Lewis rats(6-8 weeks old)by injection of complete Freund adjuvant(CFA) containing interphotoreceptor retinoid-binding protein (IRBP,1177-1191) and tuberculin (TB) into footpads,napes and back of the body,and 6 normal matched rats were used as normal controls.The diet and drinking,temperature and behavior acts of the rats were recorded during the observational duration.Ocular manifestations were examined under the slit lamp biomicroscope after modeling and scored.Eyeballs were obtained in 12 days after modeling for histopathological examination,and the ultrastructures of eyeballs were observed under the scanning electron microscope and transmission electron microscope.The use and care of the animals complied with Statement of ARVO.Results The food-intake was (190.00± 18.03)g in the model group,and that in the control group was (285.33 ±28.02) g,showing a significant difference between them (t =4.955,P =0.012).The drinking-water volume of rats was (241.67±t 18.56)ml in the model group,which was significantly less than (289.67± 18.18)ml in the control group,showing a significant difference between them (t =3.201,P =0.033).In addition,elevated temperature and tiredness were found in the model rats.Anterior chamber empyema,iris hyperemia and occlusion of the pupil appeared in the models on the 6th day and peaked on the 12nd day after immunized,with the inflammatory scores of 3.83±0.41.The infiltration of inflammatory cells were seen in anterior chamber,ciliary body and vitreous cavity under the optical microscope in the model rats.Scanning electron microscopy found uneven iris texture,coarse ciliary surface and loosen villi of retinal pigment epithelial (RPE) cells in the model rats.Under the transmission electron microscope,the infiltration of macrophagocytes on the iris,wrinkle sparse of ciliary epithelium,myeloid bodies in retinal Müller cells and vacuolus in mitochondria of RPE cells were exhibited.No obvious abnormality was found in the control rats.Conclusions Lewis rats are autoimmune status following injection of CFA with IRBP(1177-1191) and TB.The morphology and ultrastructure of eyeballs in the EAU rats can explain the finding of eyes.

OBJECTIVETo examine the effect of RhoA/Rho kinase signal pathway on TGF-beta1-induced phenotypic differentiation of human dermal fibroblasts.

METHODSThe 4th generation of primary cultured human dermal fibroblasts were stimulated with TGF-beta1, (10 ng/ml). The expression of alpha-SMA was detected after treatment with TGF-beta1, for 0, 3, 6, and 24 h. The expression of alpha-SMA was also detected after treatment with different concentration of TGF-beta1 (0, 2, 10, 50 ng/ml). Then the human dermal fibroblasts (4th generation) were stimulated with TGF-beta1, (10 ng/ml) after being treated with the RhoA/Rho kinase signaling pathway inhibitor Y-27632 (10 umol/ml). The fibroblasts were treated with nothing as sham control, or with Y-27632 (10 umol/L) only as negative control group, or with TGF-beta1 (10 ng/ml) only as positive control group. The expression of alpha-SMA was detected in all the groups. Protein expression was analyzed with ANOVA statistical method.

RESULTSalpha-SMA expression in fibroblasts with 10 ng/ml TGF-beta1 stimulation for 0, 3, 6, 24 h was 1.0, 1.9 0.2, 2.1 +/- 0. 1, 3. 1 +/- 0.1, respectively. Alpha-SMA expression in 24 h group was significantly higher than that in other three groups (n = 4, P < 0.05). alpha-SMA expression in human dermal fibroblasts after stimulation with different concentration of TGF-beta1 (0, 2, 10, 50 ng/ml) was 1.0, 1.4 +/- 0.2, 3.2 + 0.1, 3.1 +/- 0.2, respectively. alpha-SMA expression in 10 ng/ ml group was significantly higher than that in 2 ng/ml group and control group (n = 4, P < 0.05). There was no statistical difference in alpha-SMA expression between 10 ng/ml group and 50 ng/ml group (n = 4, P > 0.05). With both Y-27632 (10 micromol/L) and TGF-beta1 stimulation, the cell phenotype differentiation was inhibited. Alpha-SMA expression in experimental group (1.2 +/- 0.2) was significantly reduced, when compared with that in positive control group (2.9 +/- 0.1) (n = 5, P < 0.05). There was no significant difference (n = 5, P > 0.05) in alpha-SMA expression between control group (1.0) and negative control group (1.1 +/- 0.1).

CONCLUSIONSRhoA/Rho kinase signaling pathway should be involved in TGF-beta1-induced phenotypic differentiation of human dermal fibroblasts.

Actins ; metabolism ; Adolescent ; Cell Differentiation ; Cells, Cultured ; Fibroblasts ; cytology ; Humans ; Male ; Signal Transduction ; Skin ; cytology ; Transforming Growth Factor beta1 ; pharmacology ; rho-Associated Kinases ; metabolism ; rhoA GTP-Binding Protein ; metabolism

OBJECTIVETo establish the visual patch clamp whole-cell recording technique and study the properties and functional significance of muscarinic receptor-mediated currents in rat medial vestibular nucleus neurons (MVNn).

METHODSBrain slices containing the MVN were prepared from fifteen Wistar rats. By combining infrared differential interference contrast (IR-DIC) technique and CCD-Camera system with visual patch clamp whole-cell recording technique, twenty healthy neurons were located and muscarinic receptor-mediated currents in rat MVNn were observed and analyzed.

RESULTSVisual patch clamp technique can be used to make direct localization and to make sure of active neuron. In MVNn, a comparison of the current-voltage relationships before and during the application of muscarine, which revealed an increase in the slope of the I-V curve and the reversal potential for this response lay at (-88.4 +/- 4.9) mV (x +/- s), indicates that the activation of muscarinic cholinergic receptors leads to a decrease in potassium current. The test in the voltage sensitivity of the muscarine-induced response, which showed that the effect had a linear current-voltage relationship and reversed at (-86.7 +/- 3.5) mV, indicates that the potassium current blocked by muscarine corresponds to the voltage-insensitive leak potassium current.

CONCLUSIONSVisual patch clamp technique, which was considered better than blind patch clamp technique, can improve the success of sealing process. By the analysis of muscarinic receptor-mediated currents, the data provide support that muscarinic cholinergic mechanisms play a prominent role in the modulation of the excitability of MVNn and also offer a new idea for the efficacy of anticholinergic drugs.

Animals ; Electrophysiological Phenomena ; Neurons ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Wistar ; Receptors, Muscarinic ; physiology ; Vestibular Nuclei ; physiology

Long-segment lichen sclerosus (LS) urethral stricture is a challenge for urologists. Limited data are available for surgeons to make a surgical decision between Kulkarni and Asopa urethroplasty. In this retrospective study, we investigated the outcomes of these two procedures in patients with LS urethral stricture. Between January 2015 and December 2020, 77 patients with LS urethral stricture underwent Kulkarni and Asopa procedures for urethroplasty in the Department of Urology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine (Shanghai, China). Of the 77 patients, 42 (54.5%) underwent the Asopa procedure and 35 (45.5%) underwent the Kulkarni procedure. The overall complication rate was 34.2% in the Kulkarni group and 19.0% in the Asopa group, and no difference was observed ( P = 0.105). Among the complications, no statistical difference was observed in the incidence of urethral stricture recurrence ( P = 0.724) or glans dehiscence ( P = 0.246) except for postoperative meatus stenosis ( P = 0.020). However, the recurrence-free survival rate between the two procedures was significantly different ( P = 0.016). Cox survival analysis showed that antiplatelet/anticoagulant therapy use ( P = 0.020), diabetes ( P = 0.003), current/former smoking ( P = 0.019), coronary heart disease ( P < 0.001), and stricture length ( P = 0.028) may lead to a higher hazard ratio of complications. Even so, these two techniques can still provide acceptable results with their own advantages in the surgical treatment of LS urethral strictures. The surgical alternative should be considered comprehensively according to the patient characteristics and surgeon preferences. Moreover, our results showed that antiplatelet/anticoagulant therapy use, diabetes, coronary heart disease, current/former smoking, and stricture length may be contributing factors of complications. Therefore, patients with LS are advised to undergo early interventions for better therapeutic effects.
Male ; Humans ; Urethral Stricture/etiology* ; Retrospective Studies ; Constriction, Pathologic/surgery* ; Lichen Sclerosus et Atrophicus/surgery* ; Treatment Outcome ; Urologic Surgical Procedures, Male/methods* ; China ; Urethra/surgery* ; Postoperative Complications/etiology* ; Mouth Mucosa ; Diabetes Mellitus/etiology* ; Anticoagulants ; Coronary Disease

BACKGROUNDReports on mood regulating circuit (MRC) indicated different activities between depressed patients and healthy controls. The functional networks based on MRC have not been described in major depression disorder (MDD). Both the anterior cingulate cortex (ACC) and thalamus are all the key regions of MRC. This study was to investigate the two functional networks related to ACC and thalamus in MDD.

METHODSSixteen patients with MDD on first episode which never got any medication and sixteen matched health controls were scanned by 3.0 T functional magnetic resonance imaging (fMRI) during resting-state. The pregenual anterior cingulate cortex (pgACC) was used as seed region to construct the functional network by cortex section. The thalamus was used as seed region to construct the functional network by limbic section. Paired-t tests between-groups were performed for the seed-target correlations based on the individual fisher z-transformed correlation maps by SPM2.

RESULTSDepressed subjects exhibited significantly great functional connectivity (FC) between pgACC and the parahippocampus gyrus in one cluster (size 923) including left parahippocampus gyrus (-21, -49, 7), left parietal lobe (-3, -46, 52) and left frontal lobe (-27, -46, 28). The one cluster (size 962) of increased FC on thalamus network overlapped the precuneus near to right parietal lobe (9, -52, 46) and right cingulate gyrus (15, -43, 43) in health controls.

CONCLUSIONSAbnormal functional networks exist in earlier manifestation of MDD related to MRC by both cortex and limbic sections. The increased functional connectivity of pgACC and decreased functional connectivity of thalamus is mainly involved in bias mood processing and cognition.

Adult ; Depressive Disorder, Major ; physiopathology ; Female ; Gyrus Cinguli ; physiopathology ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Thalamus ; physiopathology

BACKGROUNDFunctional imaging studies indicate abnormal activities in cortico-limbic network in depression during either task or resting state. The present work was to explore the abnormal spontaneous activity shown with regional homogeneity (ReHo) in depression by resting-state functional magnetic resonance imaging (fMRI).

METHODSUsing fMRI, the differences of regional brain activity were measured in resting state in depressed vs. healthy participants. Sixteen participants firstly diagnosed with major depressive disorder and 16 controls were scanned during resting state. A novel method based on ReHo was used to detect spontaneous hemodynamic responses across the whole brain.

RESULTSReHo in the left thalamus, left temporal lobe, left cerebellar posterior lobe, and the bilateral occipital lobe was found to be significantly decreased in depression compared to healthy controls in resting state of depression.

CONCLUSIONSAbnormal spontaneous activity exists in the left thalamus, left temporal lobe, left cerebellar posterior lobe, and the bilateral occipital lobe. And the ReHo may be a potential reference in understanding the distinct brain activity in resting state of depression.

Adult ; Case-Control Studies ; Depressive Disorder, Major ; pathology ; Female ; Hemodynamics ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Occipital Lobe ; pathology ; Temporal Lobe ; pathology ; Thalamus ; pathology ; Young Adult

Objective To compare the differences on onset timing of acute ST segment elevation myocardial infarction (STEMI) in young and aged patients. Methods The exact onset time of symptoms was obtained from 1024 consecutive patients with STEMI admitted to our hospital between January 2000 and May 2010. Patients were classified as the middle-aged group [＜65 years old, mean (52.2 ±8.0)years, n =536]and old group [≥65 years old, (72. 2 ±5.5)years, n =488], the difference of the onset months,weeks, weekdays and hours between two groups was compared. Results The high onset timing of STEMI in middle-aged group was October and February, Friday, Saturday and Wednesday, at 10 A.m. And 10P. M. The high onset timing of STEMI in old group was October, January and March, Friday, Sunday and Monday, at 6 A.m. And 2 A. M. The incidences of STEMI in the old group were significant higher than in the middle-aged group in March ( 11.89％ ), on Sunday ( 15.97％ ) and Monday ( 17.42％ ), at 6 A.m.(6.35％)and 2 A. M. (5.74％) (all P ＜0.05) while the onset rate was significant higher in February (9. 89％ ), On Saturday16. 98％ ), At 8 P. M. (4. 86％ )and 10 P. M. (5.78％)in the middle-aged group than old group ( all P ＜ 0. 05 ). Conclusion The onset timing of STEMI in old patients was significant different from the middle-aged patients suggesting the onset timing of STEMI changes with aging.