Objective To analyze and compare the characteristics and differences of corneal endothelial cells of rhesus monkey and tree shrew eyes.Methods Corneal endothelial cells of 6 healthy rhesus monkeys (12 eyes) and 20 healthy tree shrews (40 eyes) were measured using a non-contact SP3000P specular microscope.Eight parameters were de-termined and compared with relevant parameters of human eyes reported in the literature, including minimum cell area (Smin), maximum cell area (Smax), average cell area (Savg), standard deviation of cell area (SD), coefficient of variabili-ty ( CV) , cell density ( CD) , hexagonality percentage ( HG%) and central corneal thickness ( CCT) .Results The ima-ging and measurement of all parameters could be completed in a short time both in rhesus monkeys and tree shrews.The time spent in the two kinds of animals was not significantly different.The CCT was ( 449.2 ±12.8 ) μm and ( 262.4 ± 24.6) μm, Smin was (120.4 ±26.3) S/μm2 and (153.2 ±42.9) S/μm2 , Smax was (705.0 ±130.8) S/μm2 and (468.7 ±109.3) S/μm2 , Savg was (351.1 ±26.1) and (295.4 ±18.9) S/μm2 , SSD was (113.1 ±27.4) and (75.9 ±27.3) S/μm2, CV was (31.9 ±6.0) and (25.3 ±8.3), CD was (2874.2 ±203.8) p/cell· mm-2 and (3399.3 ±224.7) p/cell· mm-2 , and the HG% was (58.6 ±9.1) and (94.0 ±9.7) in the rhesus monkeys andt tree shrews, respectively. The differences of all the above parameters between rhesus monkeys and tree shrews were statistically significant ( P<0.05 for all) .The cornea of tree shrews was significantly thinner than that of rhesus monkeys.The area and coefficient of varia-bility of tree shrews were smaller to those of rhesus monkeys, while the cell density and hexagonality percentage were higher than those of rhesus monkeys.Compared with human eyes, the CCT, CV and HG%in rhesus monkeys were highly simi-lar, while the CD was lower than that of human eyes.The CCT in tree shrew was only 60%of the rhesus monkey eyes and 50%of human eyes, while the CD and Savg were similar to that of human eyes in the 10-20 years old group.Conclu-sions The morphology and parameters of corneal endothelial cells in rhesus monkeys and tree shrews are significantly dif-ferent.There are similarities and differences among the human, rhesus monkey and tree shrew corneal endothelial cells. Both rhesus monkeys and tree shrews are appropriate experimental animals feasible for researches on human corneal endo-thelial diseases.

Near-infrared (NIR) was used as rapid analysis method to identify the sulfur fumigated Puerariae Lobatae Radix. NIR spectra of the cross-sectional and longitudinal selection of samples were acquired. Principal component analysis was conducted. The samples were randomly selected. The different pretreatment methods were compared. Discriminant models were established for every type of spectra to calculate the recognition rate. The orthogonal test and nonparametric test were used to test data normality. The result showed that absorbance values of different sections were different due to the different structure, and the raw spectra were analyzed by PCA method. The result founded that the cumulative contribution rate was arrived at 99.2% while the PC numbers were arrived at 3. The pretreatment method based on the MSC + 1D + Savitzky-Golay was the best to establish the model. For the 50 models constructed with cross-section and longitudinal spectra and total spectra, the recognition rate were (94.4 ± 0.66)%, (94.4 ± 0.66)%, (95.3 ± 0.65)%, respectively, and no difference was observed. The NIR method could be used to identify the sulfur fumigated Puerariae Lobatae Radix.
Chemistry, Pharmaceutical ; methods ; Discriminant Analysis ; Drugs, Chinese Herbal ; chemistry ; Fumigation ; Pueraria ; chemistry ; Spectroscopy, Near-Infrared ; Sulfur ; chemistry

OBJECTIVETo determine the expression of Skp2 in different prostate cancer (PCa) cell lines and tissues, and explore its influence on the androgen receptor (AR) signaling pathway and development of castration-resistant prostate cancer (CRPC).

METHODSThe expression levels of Skp2 and AR in different PCa cell lines were detected by Western blot. After knockdown of Skp2 in the C4-2 and 22RV1 cells transfected with shRNA, the expressions of AR and P27 were determined and the activity of ARR3-Luc measured by dual-luciferase reporter gene assay following treatment with dihydrotestosterone (DHT). The expressions of AR and Skp2 in human naïve PCa or CRPC specimens were detected by immunohistochemical staining followed by analysis of their differences and correlation.

RESULTSThe Skp2 protein expression level was significantly higher in the C4-2 or 22RV1 cells than in the LNCaP cells. DHT treatment increased the expression of Skp2 in the C4-2 cells, but knock-down of Skp2 significantly up-regulated the expression of the well-known downstream protein P27 and down-regulated that of AR. Consistently, DHT treatment increased the activity of ARR3-Luc, while knockdown of Skp2 remarkably decreased it in the C4-2 and 22RV1 cells (P < 0.05). In addition, significantly higher expressions of Skp2 and AR were observed in the CRPC than in the naïve specimens (P < 0.05), with a positive correlation between the two proteins (r = 0.658 1, P < 0.05).

CONCLUSIONSkp2 can enhance the expression and transcription activity of the AR protein in CRPC cells or tissues and is promising to be a critical molecular therapeutic target.

Androgens ; pharmacology ; Cell Line, Tumor ; Dihydrotestosterone ; pharmacology ; Disease Progression ; Gene Knockdown Techniques ; Humans ; Male ; Neoplasm Proteins ; genetics ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; metabolism ; Receptors, Androgen ; genetics ; metabolism ; S-Phase Kinase-Associated Proteins ; physiology ; Transcriptional Activation ; Up-Regulation

Objective To evaluate the prognostic value of urine paraquat (PQ) concentrations combined with poisoning time and creatinine clearance rate (CCr) on prognosis of patients with acute paraquat poisoning (APP). Methods A retrospective case control study was conducted. Clinical data of 96 patients with APP admitted to Department of Emergency of Shengjing Hospital of China Medical University from March 2014 to May 2016 were analyzed. The gender, age, body weight, urine PQ concentrations (determined by semi-quantitative colorimetric method), poisoning time (time from oral poison to urine detection) and CCr of patients were collected, and poisoning index (poisoning index = urine PQ concentrations × poisoning time/CCr) and simplified poisoning index (simplified poisoning index = urine PQ concentrations × poisoning time) were calculated. The patients were divided into death group and survival group according to 2-month outcome after poisoned with clinical data and telephone follow-up. The urine PQ concentrations, poisoning index, and simplified poisoning index between the two groups were compared. Binary classification logistic regression was used to analyze the risk factors affecting prognosis. Receiver-operating characteristic curve (ROC) and diagnostic test were used to analyze the prognostic value of the parameters. Results Compared with survival group, the urine PQ concentrations [mg/L: 30.00 (10.00, 100.00) vs. 10.00 (3.00, 10.00)], poisoning index [mg·h-1·μmol-1: 12.72 (1.86, 33.75) vs. 0.56 (0.18, 1.12)], and simplified poisoning index [mg·h-1·L-1: 600.00 (150.00, 1 000.00) vs. 60.00 (18.00, 120.00)] in death group were significantly increased (all P < 0.01). It was shown by logistic regression analysis that both urine PQ concentrations [odds ratio (OR) = 1.046, 95% confidence interval (95%CI) = 1.006-1.087, P = 0.022] and poisoning index (OR = 1.353, 95%CI = 0.029-1.815, P = 0.031) were independent risk factors affecting the prognosis of patients with APP. It was shown by ROC curve and diagnostic test that the poisoning index had greater area under ROC curve (AUC was 0.902) for evaluating the prognosis of patients with APP. When the best cut-off value was greater than 1.23 mg·h-1·μmol-1, the sensitivity was 90.91%, and the specificity was 73.08%. The AUC of urine PQ concentrations for evaluating the prognosis was 0.759. When the best cut-off value was greater than 20.00 mg/L, the sensitivity was 63.64%, and the specificity was 76.92%. The AUC of simplified poisoning index for evaluating the prognosis was 0.846. When the best cut-off value was greater than 135.00 mg·h-1·L-1, the sensitivity was 81.82%, and the specificity was 76.92%. Conclusion The poisoning index calculated with urine PQ concentrations combined with poisoning time and CCr has prognostic value for prognosis of APP patients, and the prognostic value of poisoning index is greater than that of the urine PQ concentrations alone.

AIM: To study the effects of taurine at different doses on renal oxidative stress and inflammation induced by paraquat in rats.METHODS: Male SD rats (n=48) were randomly divided into 4 groups: negative control group, paraquat group, paraquat + low-dose taurine group, and paraquat + high-dose taurine group.The serum levels of creatinine and urea nitrogen were detected by a biochemical analyzer.The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were measured by colorimetry.The plasma concentrations of IL-6 and intercellular adhesion molecule (ICAM)-1 were detected by ELISA.Renal reactive oxygen species (ROS) was checked by fluorescence probe dihydroethidium (DHE).The protein levels of renal p-P38 MAPK, p-ERK1/2 and p-JNK were determined by Western blot.The mRNA expression of TNF-α, IL-6 and TGF-β1 was detected by real-time PCR.RESULTS: Serum creatinine and urea nitrogen increased after paraquat poisoning, and decreased after feeding with taurine in poisoned rats, with better result in high-dose taurine group.Taurine reduced the oxidative stress and inflammation in the renal tissue, and also reduced the protein levels of p-JNK, p-ERK1/2 and p-P38 in the kidney of paraquat-poisoned rats.CONCLUSION: Taurine attenuates renal injury induced by paraquat poisoning in rats.The mechanism may be related to reducing renal MAPK activity, oxidative stress and inflammatory response.

Objective To investigate the effect of Epigallocatechin-3-gallate(EGCG)on oxidative stress and inflammatory factors in the kidney tissues of rats with paraquat poisoning,and explore the possible mechanism. Methods Rats were randomly divided into control group(group A), paraquat exposure group(group B),paraquat exposure+low dose of EGCG group(group C),and paraquat exposure+high dose of EGCG group (group D)(n=10 in each group). Kidney tissues were harvested and observed by HE staining. The level of creatinine was tested by biochemical detector. Serum oxidative stress and inflammatory factor level were detected by ELISA method. Oxidative stress in kidney tissue was determined by Western blotting and colorimetry. Inflammatory factor level in renal tissue was tested by real-time PCR. Results Kidney damage was observed in rats of groups B,C,and D. Rats in groups C and D showed less renal injury than group B,and high dose of EGCG(group D)enhanced kidney damage compared to the low dose(group C). Compared with group A,rats in the groups C and D showed lower level of 8-isoprostane,creatinine, interleukin-6(IL-6),and tumor necrosis factorα(TNF-α)in serum and malondialdehyde(MDA),3-nitrotyrosine(3-NT),and TNF-αand IL-6 mRNA in renal tissue in rats with paraquat poisoning,and group D showed the lowest mRNA level of inflammatory factor and oxidative stress. Con-clusion EGCG has protective effects on the kidney of rats with paraquat poisoning,and its mechanism may be related to the reduction of oxida-tive stress and inflammatory reaction in the kidney.

TMTP1,a 5-amino acid peptide NVVRQ,obtained by using the flagella peptide library screening in our previous studies,can be used for the labeling of malignant in situ and metastatic lesions,and even micro-metastases.In this study,TMTP1 was assessed for its ability to specifically target the malignant hematopoietic cells and metastatic lesions of hematological malignancies.FITC-TMTP 1 was chemically synthesized.Immunofluorescence assay and competitive test were carried out to determine the specific binding capacity of TMTPI to hematological malignant cell lines,including HL60,k562,SHI-1,Jurkat,Raji,El-4 and umbilical cord blood mononuclear cells.Mononuclear cells were isolated from the bone marrow of healthy subjects and patients with chronic myeloid leukemia.Then the cells were co-clutured with TMTP1 or scrambled peptides and the binding and affinity of TMTP1 peptide to the primary cells of hematological malignancies were flow cytometrically analyzed.The binding specificity of TMTP 1 to target hematological malignancies was measured in vivo by intravenous injection of FITC-conjugated TMTP1 into El-4 lymphoma-bearing mice.The results showed that TMTP1 specifically bound to the cells of a series of hematological malignancies,including HL60,k562,Jurkat,Raji,El-4 and chronic myeloid leukemia primary cells but not to bone marrow mononuclear cells from healthy subjects.By contrast,TMTP1 could bind to the metastatic foci of lymphoma originating from the EL-4 cell line while the scrambled peptide failed to do so.Moreover,the occult metastases could be identified,with high specificity,by detecting FITC-TMTP1.We are led to conclude that TMTP1,as a novel tumor-homing peptide,can serve as a marker for primary malignant and metastatic lesions for the early diagnosis of hematological malignances and a carrier of anticancer drugs for cancer treatment.

TMTP1, a 5-amino acid peptide NVVRQ, obtained by using the flagella peptide library screening in our previous studies, can be used for the labeling of malignant in situ and metastatic lesions, and even micro-metastases. In this study, TMTP1 was assessed for its ability to specifically target the malignant hematopoietic cells and metastatic lesions of hematological malignancies. FITC-TMTP1 was chemically synthesized. Immunofluorescence assay and competitive test were carried out to determine the specific binding capacity of TMTPl to hematological malignant cell lines, including HL60, k562, SHI-1, Jurkat, Raji, El-4 and umbilical cord blood mononuclear cells. Mononuclear cells were isolated from the bone marrow of healthy subjects and patients with chronic myeloid leukemia. Then the cells were co-clutured with TMTP1 or scrambled peptides and the binding and affinity of TMTP1 peptide to the primary cells of hematological malignancies were flow cytometrically analyzed. The binding specificity of TMTP1 to target hematological malignancies was measured in vivo by intravenous injection of FITC-conjugated TMTP1 into El-4 lymphoma-bearing mice. The results showed that TMTP1 specifically bound to the cells of a series of hematological malignancies, including HL60, k562, Jurkat, Raji, El-4 and chronic myeloid leukemia primary cells but not to bone marrow mononuclear cells from healthy subjects. By contrast, TMTP1 could bind to the metastatic foci of lymphoma originating from the EL-4 cell line while the scrambled peptide failed to do so. Moreover, the occult metastases could be identified, with high specificity, by detecting FITC-TMTP1. We are led to conclude that TMTP1, as a novel tumor-homing peptide, can serve as a marker for primary malignant and metastatic lesions for the early diagnosis of hematological malignances and a carrier of anticancer drugs for cancer treatment.

Objective : To observe the clearance of smear layer on the root canal wall in different action time by scanning electron microscope (SEM), and to determine the optimal amount of time using sonically activated irrigation to wash root canal in clinic. Methods: Fifty-six ex vivo human anterior teeth with single straight root canal were selected. After routine mechanical preparation, they were divided into two experimental groups according to different irrigating agents: saline group and EDTA group. Each group was assisted by VDW sonic activation EDDY. The saline group was divided into three subgroups according to the irrigating time: 5 s, 30 s and 50 s; EDTA group was divided into six subgroups according to the irrigating time: 5 s, 10 s, 20 s, 30 s, 40 s and 50 s. The control group did not undergo root canal irrigation. After irrigation, the root was cut longitudinally. The smear layer of crown, middle and apical of root canal wall was observed by SEM. Results: After irrigating for 30 seconds, there was a significant difference between the normal saline group and the control group and the 5 second group (P<0.05), and there was no difference in the middle and apical part (P>0.05). After 50 seconds, there was a significant difference in the score of the smear layer between the apical area and the other groups (P<0.05). After irrigating for 5 seconds or 10 seconds in EDTA group, there was a significant difference between the scores of the crown and middle area of the root canal and the control group (P<0.05), and there was no significant difference in the apical area (P>0.05). There was a significant difference between the 20-40 second group and the first two groups (P<0.05). There was a significant difference between the 50 second group and the other groups (P<0.05). Comparing the cleaning effect on the smear layer after 50 seconds of irrigating between the two experimental groups, the whole root canal showed significant statistical difference (P<0.05). Conclusion The EDTA-assisted sonic activated device used for 50 seconds has the best cleaning effect.

Different processed products of Coptidis Rhizoma have its unique odor, which is an important assessment index for pro- cessed products identification of Coptidis Rhizoma. Objectify odor as an entry point in this study, an electronic nose technology was used, and a suitable method for Coptidis Rhizoma measurement was built firstly. Then different processed products of Coptidis Rhizoma were detected by the method built. Finally, different processed products were identified by combining with chemometrics based on the objective odor information obtained. Electronic nose detection indicated that a significant difference in odor between different processed products was performed. Coptidis Rhizoma processed or not can be distinguished based on statistical quality control (SQC) and soft independent modeling of class analogy (SIMCA). Principle component analysis (PCA) model showed that Coptidis Rhizoma and its various processed products discriminated obviously. In addition, in order to identify the processed products of Coptidis Rhizoma, a correct recognition rate of 100% was acquired by discriminant factor analysis (DFA) , and the initial identification rate and cross-validation recognition rate of linear discriminant analysis (LDA) is 100%, 94.4% respectively. In conclusion, differentiationin odor of different processed Coptidis Rhizoma was performed by the electronic nose technology used, and different products Coptidis Rhizoma were dis- criminated by combining with chemometrics. This research can be a reference for objective identification in odor of traditional Chinese medicine, and is good for the inheritance and development of traditional experience in odor identification.
Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; Coptis ; chemistry ; Drugs, Chinese Herbal ; analysis ; Electronic Nose ; Odorants ; analysis ; Principal Component Analysis ; Rhizome ; chemistry