Objective To observe the dynamic changes of CPP32 mRNA in hippocampus following temporary whole cerebral ischemia in rats and to explore the mechanism of neuron apoptosis induced by cerebral ischemia.Methods The model of temporary whole cerebral ischemia in rats were induced by carotid artery negative pressure shunt and the expression of CPP32 mRNA were detected with fluorescence quantify RT PCR.Results Compared with the naive group, the expression of CPP32 mRNA did not increase insignificantly in sham operation group. Compared with sham operation group, the expression of CPP32 mRNA in ischemia group trended to increase at 6 h after ischemia and increased by 53% at 12 h and 223% at 24 h after ischemia. It kept in high level and decreased at 72 h after ischemia.Conclusion The expression of CPP32 mRNA in hippocampus could be induced by temporary whole cerebral ischemia. It began to increase at 6 h, reached its fastigium at 24 h and tended to decrease at 72 h after ischemia. The enhanced gene transcription of CPP32 plays an important role in the neuron apoptosis induced by cerebral ischemia.

For further improving the extraction efficiency of microwave extraction, a microwave-assisted contijuous extraction (MACE) device has been designed and utilized. By contrasting with the traditional methods, the characteristics and extraction efficiency of MACE has also been studied. The method was validated by the analysis of the triterpenoids in Ganoderma lucidum. The extraction conditions of MACE were: using 95% ethanol as solvent, microwave power 200 W and radiation time 14.5 min (5 cycles). The extraction results were subsequently compared with traditional heat reflux extraction ( HRE) , soxhlet extraction (SE), ultrasonic extraction ( UE) as well as the conventional microwave extraction (ME). For triterpenoids, the two methods based on the microwaves (ME and MACE) were in general capable of finishing the extraction in 10, 14.5 min, respectively, while other methods should consume 60 min and even more than 100 min. Additionally, ME can produce comparable extraction results as the classical HRE and higher extraction yield than both SE and UE, however, notably lower extraction yield than MASE. More importantly, the purity of the crud extract by MACE is far better than the other methods. MACE can effectively combine the advantages of microwave extraction and soxhlet extraction, thus enabling a more complete extraction of the analytes of TCMs in comparison with ME. And therefore makes the analytic result more accurate. It provides a novel, high efficient, rapid and reliable pretreatment technique for the analysis of TCMs, and it could potentially be extended to ingredient preparation or extracting techniques of TCMs.
Chemical Fractionation ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Microwaves ; Reishi ; chemistry ; Terpenes ; analysis ; isolation & purification

Objective To summarize the clinical results and safety of photodynamic therapy (PDT) through 4 years after single and multi-treatments of patients with subfoveal choroidal neovascularization (CNV) caused by age-related macular degeneration (AMD). Methods Clinical data of 73 AMD cases (95 eyes) diagnosed through fluorescein angiography (FFA), indocyanine green angiography (ICGA) and optic coherence tomography (OCT), treated with PDT were reviewed and analyzed in this hospital from June 2000 to June 2004. The changes of best corrected visual acuity (BCVA), fundus pictures, FFA, ICGA and OCT were compared before and after PDT. Follow-up time varied from 3 months to 4 years (mean, 2 years). Results The mean age of 73 patients was 67.8 years old. The BCVA was from (CF/10 cm) to 1.0. At the final follow up, the BCVA was improved (increase≥2 lines) in 39 eyes (41.1%), stabilized (?1 line) in 51 eyes (53.7%) and decreased 2 lines in 5 eyes (5.3%). Fundus hemorrhage and exudation reduced after PDT. FFA and ICGA showed CNV complete closure in 58 eyes (61.05%), partial closure in 6 eyes (6.32%), CNV incomplete closure in 22 eyes (23.16%) and recurrence in 9 eyes (9.47%). After once PDT of 12 eyes with early-stage AMD, the BCVA improved (from 0.6 to 1.5), CNV completely closed, and the OCT showed disappearance of macular edema and neursensory retinal detachment. No CNV recurred in our four years follow-up observation and the BCVA of the patients remained stable. The mean number of PDT treatment was 1.8 per eye in 95 cases. No serious local or systemic complications were encountered. Conclusions Single or multiple sessions of PDT can acheive long-term safety and efficacy. For early-stage AMD patients with minimally classic CNV, PDT can completely make CNV closed and reduce the risk of visual loss.

Purpose To evaluate short term visual acuity effects of a single photodynamic therapy(PDT) treatment with Visudyne (CIBA Vision Corp, Duluth, Ga) for choroidal neovascularization (CNV) in age related macular degeneration (AMD). Methods Definitely diagnostic AMD patients with classic CNV were treated with PDT (5 cases, 7 eyes). The data of visual acuity testing, ophthalmic examination, color photographs, optic coherence tomography, fluorescein angiograms and indocyanine green angiogram before photodynamic therapy and 1 week ,1 month after it were used to evaluate the effects of a single treatment of PDT with Visudyne. Results The visual acuity of all the treated eyes at the follow up examination at 1 month after PDT were not reduced. Distinct reduction of fluorescein leakage from CNV was noted in all patients by 1 week after PDT. Fluorescein leakage from a portion of the CNV reappeared by 1 month after treatment in 2 eyes. Conclusion PDT with Visudyne achieved short term cessation of fluorescein leakage from CNV without loss of vision or growth of classic CNV in some patients with AMD.

Objective To study the clinical results and safty of photodynamic therapy (PDT) after single and multi treatments of patients with subfoveal choroidal neovascularization (CNV) caused by wet age related macular degeneration (AMD). Methods From July, 2000 to July, 2001, 20 wet AMD patients (31 eyes) 47 88 years old (mean 68.2 years old) with best corrected visual acuity from FC/10 cm to 0.6 diagnosed through optic coherence tomography (OCT), fluorescein angiography (FFA) and indocyanine green angiography (ICGA) were treated with PDT. All cases were assigned to benzoporphyrin derivative mono acid (BPD) (6 mg per square meter of body surface area), administered via intravenous infusion of 30 ml over 10 minutes. Fifteen minutes after the start of the infusion, a laser light at 689 nm (Zeiss company, German) delivered 50 J/cm 2 at an intensity of 600 mW/cm 2 over 83 seconds on CNV. Visual acuity, photochrome of ocular fundus, OCT, FFA, ICGA were used to evaluate the effects of photodynamic therapy with BPD. Follow up of these patients was planned 1 2 week and every 3 month after PDT. Once the lesion area progressed, PDT was applied again. Tweenty cases (31 eyes) were followed up from 3 to 18 months (average 12 month).In 1 affected eye, PDT was applied fow 4 times, 4 eye for 2 times, and the other 26 eyes for 1 time. Results The visual acuity in 13 (41 9%) eyes was improved ( increase≥2 lines) after PDT. Stabilized (?1 line) in 17 (54 8%) eyes and decreased 2 lines (attributed to the recur of CNV ) in 1 (3 2%) eye. After PDT, the fundus haemorrhage and fluid leakage reduced. FFA and ICGA showed. cessation and obvious reduction of fluorescein leakage from CNV in all patients 2 weeks after photodynamic therapy, and retreatment decreased the leakage step by step. Fluorescein leakage from at least a portion of the CNV reappeared by 1 3 month after treatment in some cases. OCT also showed the reduction of the size of CNV, moreover, the edema of surrounding retina and choriodal and serous neural epithelial detachment recovered obviously. No side affect during and after PDT was noticed. Conclusions PDT with BPD can achieve short term effect on part or total cessation of fluorescein leakage from CNV without loss of vision or growth of classic CNV in patients with age related macular degeneration, retreatment of PDT was also effective.

Objective:To explore the value of pre- and postoperative red cell distribution width-to-lymphocyte count ratio (RLR) in predicting the disease-free survival (DFS) for non-metastatic colorectal cancer patients.Methods:The data of 108 patients pathologically diagnosed with non-metastatic colorectal adenocarcinoma after surgery from December 2012 to January 2020 in Hainan Hospital of PLA General Hospital were retrospectively analyzed. RLR was calculated and its differences in patients with varied clinicopathological characteristics were analyzed. The receiver operating characteristics (ROC) curve was applied to analyze the value of pre- and postoperative RLR in predicting DFS of patients, and the cut-off value was determined by Youden index. DFS of patients with different stratification stratified by a cut-off value of RLR was analyzed by using Kaplan-Meier method followed by log-rank test. Cox proportional hazards model was used to make univariate and multivariate analysis.Results:Youden index of pre- and postoperative RLR in predicting DFS was 8.86%, 9.15%; based on the above index, the patients were divided into the preoperative RLR<8.86% group (73 cases) and preoperative RLR≥8.86% group (35 cases), postoperative RLR<9.15% group (48 cases) and postoperative RLR≥9.15% group (60 cases). According to ROC curve, the area under the curve (AUC) of preoperative RLR in predicating DFS was 0.66 (95% CI 0.55-0.77, P = 0.01), and AUC of postoperative RLR in predicating DFS was 0.62 (95% CI 0.51-0.74, P = 0.04). Pre- and postoperative RLR cut-off values were used to predict 3-year DFS rate of patients with the sensitivity of 51.60% and 71.00%, respectively, and the specificity of 76.60% and 50.60%, respectively. Kaplan-Meier analysis showed that patients in preoperative RLR<8.86% group had a better DFS compared with those in preoperative RLR≥8.86% group ( χ2 = 7.35, P < 0.01); there was no statistical difference in DFS between postoperative RLR<9.15% group and postoperative RLR≥9.15% group ( χ2 = 3.69, P = 0.06). Preoperative RLR was an independent affecting factor for DFS of patients ( HR = 1.13, 95% CI 1.04-1.22, P <0.01). Conclusions:Pre- and postoperative RLR could be useful in predicating 3-year DFS for patients with non-metastatic colorectal cancer. Preoperative RLR is an independent affecting factor for DFS, and the patients with low RLR have a better prognosis.

In this study, based on web database, 324 neurotoxic compounds and 234 non-neurotoxic compounds were selected as a data set for neurotoxicity discriminative model. 6 122 molecular descriptors, including charge distribution, physicochemical and geometrical descriptors,were calculated to characterize the molecular structure of neurotoxic compounds. The combination of Cfs Subset Evaluation and Best First-D1-N5 searching was used to select molecular descriptors. A discrimination model with high accuracy was built based on the support vector machine (SVM) approach. Meanwhile, the model accuracy, sensitivity and specificity were all above 80%. Besides, 30 traditional Chinese medicine compositions with neurotoxicity were set as external validation to further verify the model accuracy,with anaccuracy of 73.333%. Using the model, 13 potential neurotoxic compounds were screened from Sophorae subprostrate Radix,4 of them were verified by literatures. The results demonstrated that the discrimination model can be applied to screen neurotoxic compounds from Chinese medicinal materials.
Computer Simulation ; Drug Evaluation, Preclinical ; methods ; Medicine, Chinese Traditional ; methods ; Models, Theoretical ; Neurotoxins ; analysis ; chemistry ; Reproducibility of Results ; Support Vector Machine

On the basis of web databases, 111 compounds with nephrotoxicity and 90 compounds without nephrotoxicity were collected as data set of nephrotoxicity discrimination model, 39 compounds with tubular necrosis and 39 compounds without tubular necrosis were collected as data set of tubular necrosis discrimination model. The 6 122 molecular descriptors, including physicochemical, charge distribution and geometrical descriptors were calculated to characterize the molecular structure of the above-mentioned compounds. CfsSubsetEval valuation method and BestFirst-D1-N5 searching method were used to select molecular descriptors. Two models with high accuracy were built based on the support vector machine (SVM) approach, respectively. Accuracy, sensitivity, specificity and matthew's correlation coefficient of the two models were all above 70%. By using 22 nephrotoxicity compounds of Chinese medicine, the nephrotoxicity discrimination model was further verified with an accuracy of 72.73%. Using the tubular necrosis discrimination model, 10 potential compounds which can cause tubular necrosis were screened from the positive results of nephrotoxicity discrimination model, 6 of them have been verified by literatures. The results demonstrated that the discrimination models can be applied to screen nephrotoxic compounds from Chinese medicinal materials, and they also offer a new research idea for the further studies on the mechanism of nephrotoxicity.
Drugs, Chinese Herbal ; toxicity ; Nephrons ; drug effects ; Support Vector Machine ; Toxicity Tests ; instrumentation ; methods

ObjectiveTo investigate the effects of prenatal taurine supplementation on the Rho-ROCK signaling pathway activity and synaptophysin (Syp) expression in brain tissues of rats with intrauterine growth restriction.MethodsEighteen pregnant Sprague-Dawley rats were randomly divided into control group, fetal growth restriction (FGR) group and taurine group, with six rats in each group. Low-protein diet was given in FGR and taurine groups to establish an FGR model. Taurine 300 mg/(kg·d) was supplemented from gestational day 12 until delivery in taurine group. The mRNA expression levels of neurite growth inhibitor-A(Nogo-A), neurite growth inhibitor receptor (NgR), Rho-A and ROCKⅡin fetal rat brain were detected using reverse transcriptase polymerase chain reaction (n=24), which are the key signaling molecules of the Rho-ROCK signal pathway. The protein expression levels of Nogo-A and NgR were detected by Western blot (n=12). The mean optical density in Nogo-A, NgR and Syp was determined by immunohistochemistry (n=18). One-way analysis of variance and LSD-t test were used for statistical analysis.Results(1) Expression of mRNA: the expression levels of Nogo-A, NgR, Rho-A and ROCKⅡ mRNA in fetal rat brain were 4.09±1.34, 3.01±0.77, 39.89±7.71 and 7.82±1.83, respectively in FGR group, and were significantly higher than in control group (1.00±0.13, 1.00±0.10, 1.02±0.30 and 1.00±0.10) (t=4.735, 5.204, 7.682 and 10.675, allP<0.05). The expressions in taurine group (1.07±0.30, 1.20±0.27, 5.36±0.41 and 1.89±0.43) were significantly lower than in FGR group (t=4.645, 4.690, 6.687 and 9.485, allP<0.05), and there was no statistical difference between taurine group and control group (allP>0.05). (2) Expression of protein by Western blot: the expressions of Nogo-A and NgR protein in fetal rat brain were 1.51±0.09 and 0.31±0.05 in FGR group, 0.82±0.06 and 0.06±0.01 in taurine group, and 1.04±0.10 and 0.09±0.12 in control group. The expression was significantly higher in FGR group than in control group (t=9.644 and 5.285, bothP<0.05). The expression was significantly lower in taurine group than in FGR group (t=14.163 and 5.825, bothP<0.05), and there was no statistical difference between taurine group and control group (allP>0.05). (3) Positive expression of protein: the positive expressions of Nogo-A and NgR protein in fetal rat brain were 0.28±0.06 and 0.11±0.02 in FGR group, 0.10±0.02 and 0.04±0.01 in taurine group, and 0.07±0.01 and 0.04±0.01 in control group. The expression was significantly higher in FGR group than in control group (t=9.778 and 7.645, bothP<0.05). The expression in taurine group was significantly lower than in FGR group (t=8.679 and 7.413, bothP<0.05), and there was no statistical difference between taurine group and control group (bothP>0.05). The positive expression of Syp protein in fetal rat brain was 0.08±0.01 in FGR group, and was significantly lower than in control group (0.16±0.04,t=4.600,P<0.05). The expression in taurine group (0.14±0.36) was significantly higher than in FGR group (t=3.181,P<0.05), and there was no statistical difference between taurine group and control group (P>0.05).ConclusionsPrenatal taurine supplementation can improve neural axon development via down-regulating the expressions of the key molecules of Rho-ROCK signal pathway in fetal rat brain tissue.

BackgroundHigh myopia is one of leading causes of blindness,so far the pathogenesis remains unclear.Two single-nucleotide polymorphisms (SNPs) of rs6885224 and rs12716080 in CTNND2 gene were recently found to be associated with high myopia in Singaporean Chinese.But whether these SNPs are related with the pathogenesis of high myopia in Han Chinese is worth studying,Objective This study was to investigate the relationship between the genetic variations of the CTNND2 gene and high myopia in Han Chinese.MethodsA case-controlled association study was designed.Nine hundred and thirty-three individuals with high myopia and 1227age- and gender-matched normal subjects were included in this study.The 5 ml of periphery blood was obtained from all subjects for the extraction of genomic DNA.The target DNA was amplified using PCR and purified by the SNaPshot method.Four SNPs rs12716080,rs917012,rs6885224 and rs16901340 in the CTNND2 gene were genotyped.This study was approved by the Ethic Committee of Sichuan Provincial People Hospital.Written informed consent was obtained from each subject before his/her enrollment.Results The frequencies of the genotypes rs6885224,rs12716080,rs917012,rs16901340 SNPs were in Hardy-Weinberg equilibrium (HWE) ( P=0.181,0.085,0.732,0.313,0.264,0.663,0.084,0.196).There were no significant differences in genotypes frequency distribution ( in turn P =0.654,0.406,0.828,0.403 ) and allele frequency distribution of the CTNND2 gene ( in turn P =0.377,0.209,0.743,0.198) between the high myopia group and normal control group.The haplotypes (TA and GA)frequencies of rs12716080 and rs917012 in the high myopia group were significantly different from those of the normal control group(TA:0.784 vs.0.719;GA:0.087 vs.0.136) (x2 =6.115,P=0.013 ;x2 =6.634,P=0.010),but those of GG were similar between the high myopia group and normal control group ( 0.123 vs.0.143,x2 =0.889,P =0.346). ConclusionsThe SNPs rs12716080,rs917012,rs6885224 and rs16901340 in CTNND2 gene were not responsible for high myopia,however,the haplotypes of rs12716080 and rs917012 are susceptible for high myopia in Han Chinese.