Objective:To investigate the protective effects of an antioxidant tert-butylhydroquinone (tBHQ) on the morphology and function of retina in early-stage experimental diabetic rats, and to explore the mechanism of its protective effect.Methods:Forty-five healthy SD rats of clean degree were randomized into normal control group, diabetes model group and tBHQ intervention group, with 15 rats in each group according to a random number table.The diabetes model was established via a single intraperitoneal injection of streptozotocin (STZ) in diabetes model group and tBHQ intervention group.Normal control group was intraperitoneally administered with an equal-volume injection of sodium citrate buffer.Rats in the tBHQ intervention group maintained a diet with 1% tBHQ for 2 weeks before the STZ injection, and the other two groups were fed with normal rat food only.Blood from tail vein was collected to assay the blood glucose level at 72 hours, 2 weeks and 4 weeks following modeling.Rat electroretinogram (ERG) was detected at 4 weeks after modeling.Morphological changes of rat retina were observed by hematoxylin and eosin staining.The apoptosis of retinal cells in different layers was detected by TUNEL assay.The expression of protein kinase B (Akt), p-Akt, endothelial nitric oxide synthase (eNOS) and p-eNOS was detected by Western blot.Müller cell line rMC-1 cells cultured in vitro were divided into 5 groups, including normal control group (72-hour culturing in normal medium), mannitol control group (72-hour culturing in medium containing 5.5 mmol/L glucose and 24.5 mmol/L mannitol), high glucose group (72-hour culturing in high-glucose medium), tBHQ intervention group (24-hour culturing in normal-glucose medium containing 5 μmol/L tBHQ, 72-hour culturing in high-glucose medium containing 5 μmol/L tBHQ), and phosphoinositide 3-kinase (PI3K) inhibitor group (6-hour culturing in normal medium containing 5 μmol/L LY294002, 24-hour culturing in normal-glucose medium containing 5 μmol/L LY294002 and 5 μmol/L tBHQ, 72-hour culturing in high-glucose medium containing 5 μmol/L LY294002 and 5 μmol/L tBHQ). The expression of Akt, p-Akt, eNOS and p-eNOS in the cells was detected by western blot.The use and care of animals complied with Regulations for the Administration of Laboratory Animals in Southwest Medical University.The study protocol was approved by the Animal Ethics Committee of Southwest Medical University (No.201711189). Results:The blood glucose level at 72 hours, 2 weeks and 4 weeks after modeling was higher in diabetic model group than tBHQ intervention group and normal control group (all at P<0.01). Four weeks after modeling, the scotopic ERG a-wave and b-wave amplitudes of diabetic model group were lower than those of normal control group and tBHQ intervention group (all at P<0.05). With edema and thickening of inner plexiform layer, thinning of inner nuclear layer and outer nuclear layer, as well as loosely arrangement and disorder of retinal layers, the number of retinal ganglion cells was decreased in diabetic model group in comparison with normal control group, all of which were improved in tBHQ intervention group in comparison with diabetic model group.There were more apoptotic retinal cells in diabetic model group than normal control group and tBHQ intervention group (both at P<0.05), which mainly existed in the outer nuclear layer.The relative expressions of p-Akt/Akt and p-eNOS/eNOS in rat retina of normal control group, diabetic model group and tBHQ intervention group were 0.76±0.11 and 0.83±0.06, 0.52±0.10 and 0.52±0.08, 1.14±0.31 and 1.03±0.13, respectively.The relative expressions of p-Akt/Akt and p-eNOS/eNOS in diabetic model group were lower than those of normal control group and tBHQ intervention group (all at P<0.01). The relative expressions of p-Akt/Akt and p-eNOS/eNOS in normal glucose group, mannitol control group, high glucose group, tBHQ intervention group and PI3K inhibitor group were 0.95±0.38 and 0.86±0.11, 0.94±0.27 and 0.74±0.29, 0.33±0.25 and 0.45±0.29, 1.32±0.37 and 1.28±0.22, 0.24±0.09 and 0.73±0.29, respectively.The relative expressions of p-Akt/Akt and p-eNOS/eNOS were significantly lower in high glucose group than those in normal glucose group and tBHQ intervention group (all at P<0.05), which were significantly lower in PI3K inhibitor group compared with tBHQ intervention group (both at P<0.01). Conclusions:tBHQ has protective effects on the morphology and function of retina in early diabetic rats, and the mechanism may be related to the activation of Akt/eNOS signaling pathway.

Diabetic retinopathy (DR) is a common microvascular complication of diabetes mellitus, with capillary occlusion, microaneurysms, and neovascularization as main pathological features, and its pathogenesis has not been fully understood.The existing clinical treatment can not cure DR, therefore, the study of the pathogenesis of DR is still a hot topic and difficult problem.Current studies have suggested that oxidative stress response is one of the important mechanisms for the development of DR, which promotes the production of excessive reactive oxygen species to induce retinal cell dysfunction and apoptosis.Imbalance of levels of nitric oxide (NO) from different types of sources in the retina under hyperglycemic stimulation also plays an important role in pathological changes, such as capillary damage, blood-retinal barrier disruption and neovascularization in DR.At the same time, NO interacts with oxides produced by oxidative stress to promote further retinal damage in DR, leading to retinal morphology abnormality and dysfunction, which ultimately cause irreversible blindness in patients.Research status of NO and oxidative stress in the pathogenesis of DR from pathophysiological changes of DR, DR and oxidative stress, DR and NO, and the role of NO in oxidative stress were reviewed in this article.

Objective: To expore the impact of teniary butyl hydroquinone (tBHQ) on Müller cells in SD rats retina under high glucose condition, and discuss the mechanism of tBHQ. Methods: The Müller cells of SD rats were cultured in vitro and the experiment was divided into normal control group, high glucose group and tBHQ intervention group.Western blot and quantitative real time PCR were used to determine the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), hemeoxygenase-1 (HO-1), hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) protein and mRNA in each group. Results: Müller cells cultured in vitro were flat with irregularly sharp.The nucleus was oval, while the cytoplasm was abundant.Adjacent cells were interwoven to a network.Western blot assay showed the overall expression of Nrf2, HO-1, HIF-1α, and VEGF in Müller cells of normal control group, high glucose group, and tBHQ intervention group were significantly different (F=73.831, 148.618, 152.269, 91.217, all at P<0.001); Among them, the relative expressions of Nrf2, HO-1, HIF-1α and VEGF proteins in the high glucose group were 0.17±0.02, 0.47±0.02, 0.67±0.07, and 0.6±0.05, which were increased in comparion with 0.06±0.01, 0.19±0.03, 0.06±0.00 and 0.07±0.02 in the normal control group, with statistically significant differences (t=4.114, 9.275, 16.479, 13.353, all at P<0.001); the relative expressions of Nrf2 and HO-1 proteins in the tBHQ intervention group were 0.40±0.06 and 0.72±0.05, which were increased by comparion with those in the higher glucose group, with statistically significant differences (t=7.847, 7.947, both at P<0.001); the relative expressions of HIF-1α and VEGF proteins in the tBHQ intervention group were 0.18±0.04, 0.26±0.07, which were decreased in comparion with those in the higher glucose group, but were increased in comparion with those in normal control group, with statistically significant differences (t=13.215, 8.444, both at P<0.001). Quantitative real time PCR showed that the relative mRNA expressions of Nrf2, HO-1, HIF-1α, and VEGF in Müller cells of normal control group, high glucose group, and tBHQ intervention group were significantly different (F=340.317, 1 582.911, 488.852, 185.699, all at P<0.001); the relative mRNA expressions of Nrf2, HO-1, HIF-1α, and VEGF proteins in the high-glucose group were 1.53±0.06, 1.50±0.04, 2.56±0.09, and 3.04±0.19, which were increased in comparion with 1.07±0.07, 0.95±0.05, 0.99±0.02, and 1.09±0.08 in the normal control group, with statistically significant differences (t=7.292, 15.014, 30.550, 18.573, all at P<0.001); The relative mRNA expressions of Nrf2 and HO-1 proteins in the tBHQ intervention group were 2.68±0.09 and 2.94±0.05, which were increased in comparion with those in the high-glucose group, with statistically significant differences (t=18.046, 39.458, both at P<0.001); The relative mRNA expression of HIF-1α and VEGF protein in the tBHQ intervention group were 1.48±0.05 and 1.6±0.08, which were decreased by comparion with those in the higher glucose group were increased in comparion with those in normal control group, with statistically significant differences (t=21.036, 13.739, both at P<0.001). Conclusions tBHQ protects Müller cells from damage in high glucose condition by activating anti-oxidative stress signaling pathway of Nrf2/ARE.

Objective To observe the effect oftert-Butylhydroquinone (tBHQ) on the expression of nuclear factor erythroid 2-related factor 2 (Nrf2),heme oxygenase (HO)-1 and phosphatidylinositol 3-kinase (PI3K) in high glucose cultured retinal Müller cells;and to investigate the anti-oxidative stress and anti-apoptotic effects oftBHQ.Methods Retinal Müller cells were divided into normal glucose group (5.5 mmol/L,N group),high glucose group (45 mmol/L,HG group) and tBHQ intervention group (HG+tBHQ group).After retinal Müller cells were cultured with high glucose for 48 hours,the pretreatment with tBHQ (20 μmol/L) induced the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and HO-1.The Müller cells were identified by immunofluorescence staining.The expressions of Nrf2,HO-1,PI3K,B-cell lymphoma-2 (Bcl-2) and Bax were detected by Western blot and real-time fluorescence quantitative PCR.Flow cytometry was used to detect the apoptosis of retinal Müller cells in rats.Results Müller cytoplasm and nucleus GS showed strong positive,large cell body,abundant cytoplasm,uniform green fluorescence;nuclear DAPI staining round or oval,clear boundary.The expression of Nrf2 protein (t=4.114,P=0.006),HO-1 protein (t=9.275,P=0.000),Nrf2 mRNA (t=7.292,P=0.000) and HO-1 mRNA (t=15.014,P=0.000) in the HG group were higher than those in the N group.The expressions of Nrf2 protein (t=7.847,P=0.000),HO-1 protein (t=7.947,P=0.000),PI3K protein (t=5.397,P=0.002),Bcl-2 protein (t=6.825,P=0.000),Nrf2 mRNA (t=18.046,P=0.000),HO-1 mRNA (t=39.458,P=0.000),PI3K mRNA (t=4.979,P=0.003) and Bcl-2 mRNA (t=9.535,P=0.000) in the HG+tBHQ group were significantly higher than those in the HG group.The protein and mRNA expressions of Bax protein in the HG+tBHQ group were significantly lower than those in the HG group (t=14.998,16.520;P=0.000,0.000).Flow cytometry showed that the apoptosis rate of Müiller cells in the HG group was significantly higher than that in the N group (t=39.905,P=0.000).The apoptosis rate of Müller cells in the HG+tBHQ group was significantly lower than that in the HG group (t=21.083,P=0.000).Conclusion tBHQ can inhibit the apoptosis of retinal Müller cells by up-regulating the expression ofNrf2,HO-1 and PI3K.

Objective To investigate the epidemiological characteristics of cerebral palsy(CP)in children aged 1-6 years in China,including the incidence,prevalence,type of CP,etiology,prevention and rehabilitation status. Methods The survey was carried out by standard questionnaires,multi-center collaboration,stratified-cluster ran-dom sampling method.The surveyed adopted the following principles:streets in the city and villages in the rural areas, and the number of the urban and rural children was the same,and the proportion of children in each age group was balanced.The investigation areas included provinces and autonomous regions,including Heilongjiang,Beijing,Henan, Shandong,Shanxi,Shaanxi,Anhui,Hunan,Guangxi,Guangdong,Chongqing and Qinghai,and 323 858 children were in-vestigated.Results The incidence of CP was 2.48‰(155/62 591 cases),and the prevalence was 2.46‰(797/323 858 cases)(1-6 years old).The prevalence varied in different regions,in which the highest prevalence was 5. 40‰(54/9 998 cases)in Qinghai province,and the lowest prevalence was 1.04‰(47/45 133 cases)in Shandong province.The prevalence of the males(2.64‰,461/174 391 cases)was higher than that of the females(2.25‰, 336/149 467 cases),and the difference was statistically significant(P<0.05).The types of CP were spastic type (58.85%,469/797 cases),mixed type(13.17%,105/797 cases),dyskinetic(9.79%,78/797 cases),hypotonic (8.28%,66/797 cases),ataxia(6.25%,52/797 cases)and rigid(3.39%,27/797 cases)respectively in 797 CP children.The first three risk factors for CP were long -term exposure to harmful physical factors during pregnancy, whether there were birth defects among the three generations of relatives of the children,such as children's peers, parents or grandparents,whether there were birth defects among the children's peers,parents or grandparents,and neonatal jaundice or persistent jaundice.Among 797 CP children,79.67% of the children with CP were timely detected and treated in the local hospitals,while the other 19.93% of them were not timely treated.The places which could give them timely detection and early diagnosis and treatment were general hospitals(42.97%),Maternity and Infant Hospitals (27.03%)and Children's Hospitals(20.31%). The main rehabilitation methods for 797 children with CP were 34.58% in the hospitals or rehabilitation centers,31.61% in the communities(including at home),33.80% mainly in the medical institution,and in the communities they could also receive partially rehabilitation services. Conclusions The prevalence of CP in China is coincident with international levels.The prevalence rate of CP in males is higher than that in females.The types of CP distribution are accorded with international distribution characteristics.There were still some children with CP who could not receive timely detection and treatment.Rehabilitation at the medical institutions is the chief way and proper rehabilitation guidance should be carried out in the communities.

Objective To observe the clinical effect of arthroscopic surgery on patients with rotator cuff tear injury. Methods One hundred and twenty patients with rotator cuff tears admitted to the Department of Orthopaedic of the First Hospital of Changsha from January 2013 to September 2015 were enrolled. Sixty patients treated with traditional conservative methods were assigned in a control group, the whole arthroscope or shoulder arthroscope was used to assist performing microsurgical incisions for another such 60 patients in the observation group. Visual analogue scale (VAS) of pain was applied to evaluate the pain of the patients with rotator cuff injury before and after treatment in both groups. The function of the shoulder joints and recovery situation were evaluated by using the shoulder functional system score and the American shoulder and elbow surgeon score (ASES). Results The results showed that all patients had no postoperative complications such as secondary fracture of the rotator cuff tear, infection, incision non-healing, etc. Compared with those before treatment, in both groups, the VAS scores were obviously lowered after treatment, while ASES and shoulder function system scores after treatment were significantly higher than those before treatment, and the changes in the observation group were more pronounced than those in the control group (VAS: 0.82±0.11 vs. 2.20±0.59, ASES: 82.21±10.81 vs. 70.53±6.21, pain score: 9.81±0.21 vs. 9.11±0.51, function score: 9.70±0.09 vs. 8.80±0.40, anterior flexion score: 4.59±0.41 vs. 4.20±0.61, all P ＜ 0.05). Conclusion The clinical effect of arthroscopic surgical treatment of patients with rotator cuff tear injury is relatively satisfactory and worthy to be deeply studied clinically.

Objective: To investigate the effect of hypertensive disorder complicating pregnancy (HDCP) on the mortality and early complications of premature infants. Methods: The general clinical data of preterm infants with gestational age 24-36^{+ 6} weeks were collected from the cooperative units in the task group from January 1, 2013 to December 31, 2014.According to the severity of HDCP, the infants were divided into 4 groups: HDCP group, preeclampsia group, eclampsia group and non HDCP group, the mortality and major complications of preterm infants were compared, and the influencing factors were analyzed. Results: The mortality rate of preterm in the HDCP group was significantly higher than that of non HDCP group, and there was statistical significance (χ²=9.970, P=0.019). Eclampsia had a highest fatality rate (4.8%) in the early stage, compared with non HDCP group (2.2%), and the difference was statistically significant.Comparison of HDCP group (1.8%) and eclampsia group (3.2%) suggested that there was no statistically significant difference.The incidence of respiratory distress syndrome (RDS) in preterm in HDCP group was significantly higher than that of non HDCP group, and there was statistical significance (χ²=13.241, P=0.004). Eclampsia group showed the highest incidence (35.4%), compared with non HDCP group (16.2%), the difference was statistically significant, but compared with HDCP group (19.9%), preeclampsia group (17.1%), there was no significant diffe-rence.The incidence of bronchopulmonary dysplasia (BPD) in preterm in HDCP group was significantly higher than that of non HDCP group (χ²=9.592, P=0.022), the highest incidence showed up in eclampsia group (9.7%), compared with non HDCP group (2.0%) and HDCP group (1.7%), the difference was statistically significant.But there was no statistically significant difference, compared with preeclampsia group.As the degree of HDCP aggravated, the incidence of BPD gradually rose.There was no significant impact on necrotizing enterocolitis (NEC), retinopathy of prematurity (ROP), intraventricular hemorrhage (IVH) and sepsis of HDCP (χ²=7.054, 7.214, 0.358, 3.852; P=0.070, 0.065, 0.949, 0.278). Considering the overall outcome of the child, that was, whether the child died or survived, he had at least one complication, and HDCP had an effect on it (χ²=15.697, P=0.001), so the incidence increased while the degree of HDCP rose gradually.After adjusting gestational age, birth weight, sex, way of delivery, placental abruption and front placenta, prenatal hormonal, gestational diabetes, neonatal asphyxia and other factors, the results displayed that HDCP was the factor leading to the death of premature baby (OR=2.159, 95%CI: 1.093-4.266), and comparison between preeclampsia and eclampsia showed no statistical difference (P=0.714, 0.389); HDCP had no significant influence on RDS, BDP, ICH, NEC, ROP and sepsis. Conclusions HDCP leads to increased risk of premature death, but also leads to the increased incidence of RDS and BPD, but it had no obvious effect on NEC, ROP, IVH, sepsis and other complications.

Objective To investigate the cellular viability and mitochondrial reactive oxygen species (ROS) production of the Müller cells under high glucose condition,and explore the protection role of the 5,6-dihydrocyclopenta-1,2-dithiole-3-thione (CPDT) on Müller cells.Methods Müller cells from Sprague Dawley rats were divided into 5 groups randomly,including 25 mmol/L normal glucose group (group A) and 65 mmol/L high glucose group (group B).High glucose group with 45,60,70 μmol/L CPDT and cultured them 72 hour was set as group C,D and E.Water soluble tetrazolium salt (WST)-8 was used to measure the cellular viability.Flow cytometry was used to measure the active oxygen and apoptosis index.The expression of nuclear factor erythroid 2-related factor 2 (Nrf2),hemeoxygenase-1 (HO-1),Bcl-2 and Bax protein were measured by Western blot.Results Compared with group A,the WST-8 showed that the viability of Müller cells apparently decreased in group B (t=39.59,P＜0.05).Compared with the group B,the viability of Müller cells had changes in group C (t=0.97,P＞0.05),but recovered in group D and E (t=-4.17,-7.52;P＜0.05).Compared with group A,the FCM showed that the mitochondrial ROS levels was higher in group B (t=-30.99,P＜0.05).Compared with group B,the mitochondrial ROS levels were decreased in group D (t=27.68,P＜0.05).Compared with group A,Bax,Nrf2 and HO-1 increased (t=-11.03,-63.17,-11.44;P＜0.05),while the bcl-2 decreased in group B (t=7.861,P＜0.05).Compared with the group B,Nrf2,HO-1 and Bax decreased (t=15.11,26.59,6.27;P＜0.05),while the bcl-2 increased in group D (t=-6.53,P＜0.05).Conclusions Under the high glucose,CPDT may reduce the mitochondrial ROS levels and the expression of Nrf2,HO-1 and Bax protein of Müller cells.It may inhibit apoptosis through activating the Nrf2/HO-1 pathway and balancing of level of Bcl-2 protein and mitochondrial ROS.

Objective To investigate association between polymorphisms of the tumor necrosis factor α-induced protein 3 (TNFAIP3)interacting protein 1 (TNIP1)gene and systemic lupus erythematosus (SLE)in a Chinese Han population. Methods Blood samples were collected from 284 patients with SLE of Han nationality(SLE group)and 630 healthy controls of Han nationality (control group). Ligase detection reaction (LDR)was performed to determine the genotypes of 120 single nucleotide polymorphisms (SNPs)in the TNIP1 gene. Data were analyzed with the PLINK 1.07 package and Haploview software. Results After quality control, data on 105 SNPs underwent statistical analysis finally. Four SNPs including rs3805433, rs12516176, rs6869605 and rs4958882 in the TNIP1 gene were significantly associated with SLE (OR: 1.50 - 1.53, all P < 4.72 × 104), and there was a significant increase in the frequency of rs3805433 C, rs12516176 C, rs6869605 C and rs4958882 G alleles in the SLE group (0.301 - 0.306)compared with the control group (0.221 - 0.225). There was strong linkage disequilibrium between these 4 SNPs (r2 ≥ 0.871, D′ ≥ 0.938). In addition, moderate linkage disequilibrium was observed between these 4 SNPs and a previously reported SLE-related SNP rs10036748 (r2 ≥ 0.073, D′ ≥ 0.868). The frequency of the haplotype H2: CCCGT was significantly higher in the SLE group than in the control group(0.290 vs. 0.210, OR = 1.54, P < 4.72 × 10-4). Conclusion TNIP1 gene polymorphisms are associated with SLE in Chinese Han population.

Objective To explore the efficacy of constructing the neourethra using a bladder anterior wall for the treatment of female total urethral stricture or atresia.Methods We retrospectively reviewed 11 female patients with total urethral stricture or oblitalition,who were underwent a procedure of reconstructive neourethra using a bladder anterior wall,from January 2009 to November 2015.Of the 11 patients,urethral stricture was associated with vesicovaginal fistula and a severe hydrocolpos in the proximal vagina because of vaginal anterior strictures or atresia in four girls.The mean age was 16 years (ranging 5-48 years) in all patients.The etiology was posttraumatic urethral injuries after pelvic fracture in 9 patients,radical urethral resection because of urethral cancer in 1 patient and congenital bladder exstrophy with an absent urethra in 1 patient.All patients underwent a procedure of neourethral construction under general anesthesia.The bladder anterior wall,which was about 2.0 to 2.5 cm in width and 4.0 ～4.5cm in length,was separated from bladder neck to middle partion of the anterior bladder wall.The bladder flap was tubularized around a 12-14 French catheter using continuous 4-0 polyglycolic acid sutures for the mucosa and interrupted sutures of 3-0 polyglycolic acid for the muscle.The tubularized flap was then flipped caudally to the site of the original external urethral meatus to form a new urethra.4 patients with severe stenosis or oblitalition of the distal vagina underwent a procedure of vaginoplasty at same time,including island vulvar flaps enlarging vaginoplasty in two girls and reconstructive vaginal orifice using the proximal enlargedvagina wall in other two girls.Results There were no serious complications postoperatively.The catheter was removed 3 ～4 weeks after the operation.7 patients were completely continent with excellent voiding,3 patients had stress incontinence.One patient experienced dysuria.And the urethroscopy in this case showed that the mucosal prolapse was present at the 12 to 3 o'clock position on the neck of the bladder,which caused urinary obstruction.Endoscopic resection of the prolapsed mucosa was performed.The patient could easily void without incontinence after the operation.The patients were followed up a median of 38 months,(ranging 6-72 months).2 patients experienced dysuria 3 and 4 months after operation,separatively.Examination showed that the mucosal prolapse was present at the position on the neck of the bladder in one patient and urethral meatal stenosis in another patient.The two patients were separatively underwent a procedure of endoscopic resection of the prolapsed mucosa and meatal urethroplasty,using vulvar flap.All of them could easily void without incontinence after the operation.Of the 3 patients with stress urinary incontinence,one underwent a procedure of TVT-O one year later,and after which continence was achieved with good voiding;the other two cases were awaiting for reoperation.Four cases of postoperative vaginal fluid disappeared with unobstructed micturition.Conclusions Female neo-urethral reconstruction using the bladder anterior wall flap was a reliable technique for the management of complete urethral stricture or obliteration.