Objective To investigate the polymorphism of the vitamin D receptor gene (VDR)TaqⅠin relation to diabetic retinopathy. Method Fragment length discrepant allele specific PCR(FLDAS-PCR) were used to determine VDR genetypes in 158 patients with diabetic retinopathy and in 198 normal subjects. Results The frequency distribution of VDR genotypes in diabetic retinopathy patients was 106 (67.1%) in TT, 33(20.9%) in Tt, 19(12.0%) in tt; and in normal persons was 165 (83.3%) in TT, 23(11.6%) in Tt, 10 (5.1%) in tt. There was a significant difference between diabetic retinopathy patients and normal persons in distribution of VDR gene TaqⅠgenotypes(Plt;0.05). Conclusions There is some distribution alterations of VDR gene polymorphism in diabetic retinopathy patients. (Chin J Ocul Fundus Dis, 2006, 22: 94-96)
目的 探讨聚乙二醇滴眼液对准分子激光上皮下角膜磨镶术（LASEK）后角膜雾状混浊（haze）影响的临床研究。 方法 回顾性分析2009年3月－2010年3月行LASEK的患者45例90只眼的临床资料。患者屈光度（?4.68 ± 2.30）D（?1.75～?6.50 D），随机分为聚乙二醇滴眼液组（试验组）和对照组，术后定期随访观察两组角膜haze形成、角膜上皮愈合时间、视力恢复情况及屈光度。 结果 术后1、3、6个月，两组haze形成差异无统计学意义（P＞0.05），试验组术后第1、3、5天角膜刺激症状明显轻于对照组（P＜0.05），上皮愈合速度较对照组更快（P＜0.05）。 结论 LASEK术后应用聚乙二醇滴眼液虽然能减轻术后患者角膜刺激症状,能促进角膜上皮更快愈合，但是不能减轻haze的形成。
This study aimed to assess the therapeutic effect of mannital administration on acute hemorrhagic necrotizing pancreatitis (AHNP), 28 Wistar rats were randomily divided into therapeutic group and control group after induction of AHNP by retrograde intraductal injection of 5 percent sodium taurocholate. The rats of therapeutic group received intravenous 20% mannital (1g/kg) through tail vein, once in 12 hours, until the end of experiment; control group received saline (5.0 ml/kg) with the same way. Blood of all the rats were collected from heart and the rats were killed after 96 hours. Results: lipid peroxide (LPO) in pancreatic tissue, LPO in serum, lactate dehydrogenase (LDH), alpha-1-antitrypsin (α1-AT), glutamicoxalacetic transaminase (GOT), necrotizing square of pancreatic tissue in the therapeutic group were significantly less than those in control group (Ｐ＜0.05 or Ｐ＜0.01). The damage to pancrease, heart, liver, kidney in the therapeutic group were lighter than those of the control group and the mortality was lower (Ｐ＜0.05).Conclusions: Mannital can scavenge the oxygenderived free radicals and play a therapeutic role in AHNP.
【摘要】 目的 观察在不同剂量乙醇作用下大鼠下丘脑和脊髓神经细胞P物质的表达情况和扫描电子显微镜（SEM）下神经细胞的形态学变化，探讨乙醇作用下大鼠行为学改变的相关机制。 方法 通过福尔马林实验观察大鼠在不同剂量乙醇及时间作用下行为学的改变；采用免疫组织化学技术检测不同剂量乙醇作用下大鼠脊髓和下丘脑神经细胞中P物质的表达，通过扫描电子显微镜观察神经细胞的形态学变化。 结果 乙醇灌胃后0～2 h大鼠舔足次数有不同程度的变化，组间比较差异有统计学意义（Plt;0.05），灌胃2 h大鼠下丘脑和脊髓P物质表达程度与乙醇剂量有相关关系，扫描电子显微镜下各组大鼠的神经细胞形态学变化显著。 结论 急性乙醇中毒可引起大鼠对疼痛反应的变化，其程度与乙醇剂量和作用时间有关，大鼠下丘脑和脊髓神经细胞中P物质的表达强度与乙醇剂量和作用时间有关。【Abstract】 Objective To observe the expression of substance P(SP)in the hypothalamus and spinal cord nerve cells of rats with different concentrations of ethanol, and to observe the morphological changes of nerve cells by scanning electron microscopic(SEM) for elucidating the mechanism of ethological changes effected with ethanol. Methods Ethological changes were detected through the formalin test; SP expressions in the hypothalamus and the spinal cord were evaluated with immunohistochemistry technology, and the morphological changes of nerve cells were observed by SEM. Results The frequency of licking foot changed when the rats were gavaged with different concentrations of ethanol among zero to two hours, the difference between two groups was statistical signifcant (Plt;0.05). The expression level of SP and the morphological changes of nerve cells in hypothalamus and spinal cord had relationship with the ethanol concentration. Conclusions Acute alcoholism could cause pain dysfunction in rats. The frequency of licking foot of rats is correlated to the role of the time closely. The expression intensity of SP in the hypothalamus and the spinal cord nerve cells are correlated to the concentration of ethanol closely.
ObjectiveTo verify the expression change of insulin-like growth factor-Ⅰ (IGF-Ⅰ) protein and its mRNA before and after Roux-en-Y gastric bypass surgery (RYGB) in obese rats, and to investigate the relationship between the expression of IGF-Ⅰ and proliferation/apoptosis of adipose cells. Methods① Seventy male SD rats were raised at the SPF level circumstance and were randomly divided into control group (NC group, 10 rats) and high fat diet group (60 rats). Rats of high fat diet group were given specific high fat formula diet, rats of NC group were given particular formula diet. After 6 weeks, the body weights of the rats in high fat diet group were measured, and the 20 rats of top weight were selected. The 20 obese rats were randomly divided into 2 groups:gastric bypass (GB) group (n=10) and sham-operation group (SO group, n=10). RYGB were administered to the rats of GB group, and for rats of SO group, sham operations were performed. Rats of NC group did not receive any surgery. Inguinal adipose tissues[represented the subcutaneous adipose tissue (SAT)] and epididymal adipose tissues[on behalf of visceral adipose tissue (VAT)] were taken during operation in rats of GB group and SO group respectively (0.5 g), and 12 weeks after operation in all rats of three groups. The expressions of IGF-Ⅰ protein and its mRNA in adipose tissue were detected by Western blot and real-time fluorescence quantitative PCR. ② Transfection experiment. SAT cells were divided into blank control group (BC group, without transfection), IGF-Ⅰ(+) group (gene overexpression group), IGF-Ⅰ(+) empty vector group, IGF-Ⅰ(-) group (gene silencing group), and IGF-Ⅰ(-) empty vector group. Cells were transfected with corresponding vectors with 3 duplicated holes of each group. Cell viability and apoptosis assays were carried out in 48 hours after transfection. Expressions of protein kinase B (AKT), phosphorylated protein kinase B (p-AKT), phosphoinositide 3-kinase (PI3K), and phosphorylated phosphoinositide 3-kinase (p-PI3K) were detected by Western blot meanwhile. ③ Wortmannin experiment. SAT cells were divided into Wortmannin (+) IGF-Ⅰ(+) group, Wortmannin (+) IGF-Ⅰ(-) group, Wortmannin (-) IGF-Ⅰ(+) group, and Wortmannin (-) IGF-Ⅰ(-) group, which were transfected with corresponding vectors for 24 hours, then adding Wortmannin (0.1 mmol/L). After 24 hours, the expression levels of AKT, p-AKT, p-PI3K, PI3K, and GAPDH were detected by Western blot. Results① PCR results showed that, in SAT, compared with preoperative GB group, the expression levels of IGF-Ⅰ mRNA and its protein in postoperative GB group were both lower (P < 0.01). However, the expression levels of IGF-Ⅰ mRNA and its protein between preoperative SO group and postoperative SO group showed no significant difference (P > 0.05). In VAT, the expression levels of IGF-Ⅰ mRNA and its protein in 5 groups showed no significant difference (P > 0.05). ② The MTT results showed that, IGF-Ⅰ(+) group harbored stronger proliferation abilities compared with its negative control group (P=0.04), whereas IGF-Ⅰ(-) group had lower abilities compared with its negative control group (P=0.04). The results of flow cytometry assay showed that, the apoptosis rate of IGF-Ⅰ(+) group was lower (P=0.04) than that of the corresponding negative control group, and it was higher in IGF-Ⅰ(-) group than that of the corresponding negative control group (P=0.04). ③ Compared with IGF-Ⅰ(+) empty vector group, p-PI3K/PI3K ratio (P=0.03) and p-AKT/AKT (P=0.04) ratio of IGF-Ⅰ(+) group were increased; compared with IGF-Ⅰ(-) empty vector group, p-PI3K/PI3K ratio (P=0.04) and p-AKT/AKT ratio (P=0.04) of IGF-Ⅰ(-) group were decreased. The p-AKT/AKT ratio of Wortmannin (-) IGF-Ⅰ(+) group was higher (P < 0.05) than that of Wortmannin (+) IGF-Ⅰ(+) group; the p-AKT/AKT ratio of Wortmannin (-) IGF-Ⅰ(-) group was lower than that of Wortmannin (-) IGF-Ⅰ(+) group (P < 0.05). ConclusionsIGF-Ⅰ is involved in the accumulation of subcutaneous fat in rats. RYGB can significantly reduce the expression levels of IGF-Ⅰ mRNA and its protein in subcutaneous fat of rats, so as to achieve the effect of weight loss.
ObjectiveTo observe the clinical features and visual prognostic factors of ethambutol-induced optic neuropathy (EON).MethodsA cohort study. Twenty-four inpatients (46 eyes) identified as EON in Neuro-Ophthalmology Department of Chinese PLA General Hospital from January 2014 to December 2017 were enrolled, including 14 males (26 eyes) and 10 females (20 eyes) with a ratio of 1.4/1 male/female. The average age was 42.79±15.12 years and the average weight was 62.46±12.31 kg. The average time duration between oral administration of ethambutol and occurrence of EON was 9.94±16.49 months. The average time of ethambutol duration was 7.06±11.68 months, with an average accumulative dose of 156.7±1 779.0 g and the average daily dose of 15.07±8.95 mg/(kg·d). All patients were tested with visual acuity, fundus photos, colour vision, OCT, visual field, VEP, orbital MRI and the gene of OPA1 and mitochondrial deoxyribonucleic acid (mtDNA). All the patients accepted drug withdrawal immediately after diagnosis, and were given the treatment of systemic nerve nutrition and improvement of microcirculation for 2 weeks. The time of follow-up was more than 12 months. According to whether the visual acuity (VA) in any of eyes was over than 0.1 at the last follow-up, all the patients were divided into two groups: the bad VA group (VA less than or equal to 0.1) and the better VA group (VA over than 0.1) group. The χ2 test and Fisher's exact probabilistic method test were used to compare the counting data between groups, and the Wlincox rank sum test was used to compare the measurement data. Multiple factors of VA outcome between the patients with bad or better va were analyzed by logistic regression.ResultsThirty eyes (65.2%) had VA less than or equal to 0.1 and 5 eyes (10.9%) had VA over than 0.5 at EON onset. The VA of the rest 11 eyes (23.9%) was higher than 0.1 and lower than 0.5. At the last follow-up, 20 eyes (43.5%) had VA less than or equal to 0.1 and 9 eyes (19.6%) had VA over than 0.5, the VA of the rest 17 eyes (36.9%) was higher than 0.1 and lower than 0.5. Fundus examination revealed 7 eyes (15.3%) with optic disc edema. OCT revealed significant loss of the retinal nerve fiber layer (RNFL) in the affected eyes, mainly in the temporal RNFL of the optic disc. All patients had dyschromasia, mainly in distinguishing the color of red and green. The types of visual field defect was as following: central dark spot (52.2%), diffuse visual acuity decreased (30.4%), temporal hemianopsia (17.4%). Orbital MRI revealed that 12/24 (50.0%) patients had T2 lesions with T1 enhancement in 6/24 patients (25.0%). Genetic test showed that 4 patients (16.7%) had gene mutation. Among them, there were 2 patients with OPA1 mutation, 1 with mtDNA 14340 point mutation and 1 with the mtDNA 11778 point mutation. Thirteen patients showed better VA outcomes (over than 0.1) while 11 showed bad VA outcomes after discontinuation of ethambutol. Between the better VA group and the bad VA group, there were statistically significant differences in the daily dose of ethambutol and gene mutation (P=0.031, 0.023). The daily dose was related to visual prognosis of EON while only the daily dose of more than 18 mg/(kg·d) may lead to bad VA outcomes according to the logistic analysis (95% CI 0.007-0.736, OR=0.069, P=0.027).ConclusionsEON may have OPA1 and mtDNA mutation with more bilateral eyes involved and less optic edema, which about 43.5% of the patients showed irreversible visual impact. The daily dose of ethambutol is related to the vision recovery.
Objective To investigate the cellular compatibil ity of polyvinyl alcohol (PVA)/wild antheraea pernyisilk fibroin (WSF), and to explore the feasibil ity for tendon tissue engineering scaffold in vitro. Methods The solutions of WSF (11%), PVA (11%), and PVA/WSF (11%) were prepared with 98% formic acid (mass fraction) at a mass ratio of 9 : 1. The electrospinning membranes of WSF, PVA, and PVA/WSF were prepared by electrostatic spinning apparatus. The morphologies of scaffolds were evaluated using scanning electronic microscope (SEM). The tendon cells were isolated from tail tendon of 3-dayold Sprague Dawley rats in vitro. The experiment was performed using the 3rd generation cells. The tendon cells (1 × 106/mL) were cocultured with PVA and PVA/WSF electrospinning film, respectively, and MTT test was used to assess the cell adhesion rate 4, 12 hours after coculture. The tendon cells were cultured in PVA and PVA/WSF extraction medium of different concentration (1, 1/2, and 1/4), respectively; and the absorbance (A) values were detected at 1, 3, 5, and 7 days to evaluate the cytotoxicity. The composite of tendon cells and the PVA or PVA/WSF scaffold were observed by HE staining at 7 days and characterized by SEM at 1,3, 5, and 7 days. Results The solution of WSF could not be used to electrospin; and the solution of PVA and PVA/WSF could be electrospun. After coculture of tendon and PVA or PVA/WSF electrospinning membranes, the cell adhesion rates were 26.9% ±0.4% and 87.0% ± 1.0%, respectively for 4 hours, showing significant difference (t=100.400, P=0.000); the cell adhesion rates were 35.2% ± 0.6% and 110.0% ± 1.7%, respectively for 12 hours, showing significant difference (t=42.500, P=0.000). The cytotoxicity of PVA/WSF was less significantly than that of PVA (P lt; 0.05) and significant difference was observed between 1/2 PVA and 1/4PVA (P lt; 0.05). HE staining and SEM images showed that the tendon cells could adhere to PVA and PVA/WSF scaffolds, but that the cells grew better in PVA/WSF scaffold than in PVA scaffold in vitro. Conclusion PVA/WSF electrospinning membrane scaffold has good cell compatibility, and it is expected to be an ideal scaffold of tendon tissue engineering.