Background Neurotransmitter secretion disorder induced by chronic manganese poisoning has always been one of the important causes of body injury, but the mechanism of neurotransmitter secretion disorder caused by manganese is not clear at present. Objective To investigate the effects of presynaptic membrane intracellular protein 13-1 (Munc13-1) and synapse fusion protein binding protein 18-1 (Munc18-1) on dopamine secretion dysfunction induced by manganese chloride (MnCl₂) in human neuroblastoma (SH-SY5Y) cells. Methods A SH-SY5Y cell model induced by MnCl₂ was established. Cell viability was measured by MTT assay. Four experimental groups were set up: control group and low-, medium-, and high-dose manganese groups (0, 100, 200, and 400 μmol·L⁻¹ MnCl₂). They were treated with corresponding doses of MnCl₂ for 24 h. The secretion of dopamine was measured by enzyme-linked immunosorbent assay. The mRNA expression of Syntaxin-1 was detected by real-time quantitaive PCR. Total cell proteins were extracted, and the protein expression levels of Munc13-1, Munc18-1, and Syntaxin-1 were detected by Western blotting. The correlations of MnCl₂ exposure and dopamine secretion with the protein expressions of Munc13-1 and Munc18-1 were also analyzed by Pearson correlation. Results Compared with the control group, the cell viability rate decreased gradually with the increase of manganese exposure concentration, and the difference between the medium- and the high-dose manganese groups was statistically significant (P<0.05). The concentration of dopamine in cell culture medium of all manganese exposure groups decreased with the increase of manganese concentration, and compared with the control group and the low-dose manganese group, the medium- and the high-dose manganese groups were statistically significant (P<0.05). The expression of Syntaxin-1 at mRNA or protein level did not change significantly among groups (P>0.05). Compared with the control group, the protein expression of Munc13-1 decreased and that of Munc18-1 increased with the increase of manganese concentration (P<0.05). Compared with the low-dose manganese group, the changes of Munc13-1 protein in the high-dose manganese group and Munc18-1 protein in the medium- and high-dose manganese groups had statistical significance (P<0.05). Compared with the medium-dose manganese group, the protein changes of Munc18-1 in the high-dose manganese group were statistically significant (P<0.05). The correlation analysis showed that MnCl₂ dose was negatively correlated with Munc13-1 protein expression (r=−0.898, P<0.05), and positively correlated with Munc18-1 protein expression (r=0.678, P<0.05). Dopamine secretion was positively correlated with Munc13-1 protein expression (r=0.932, P<0.05), and negatively correlated with Munc18-1 protein expression (r=−0.817, P<0.05). Conclusion The inhibition of dopamine secretion in SH-SY5Y cells induced by manganese exposure is related to up-regulation of Munc18-1 and down-regulation of Munc13-1 expression levels, which may be one of the reasons for nerve injury caused by manganese.

The HPLC fingerprint determination method of Jinzhen oral solution was established to provide a new method for quality control of Jinzhen oral solution. RP-HPLC was used for phenomenex Luna C18 (4.6 mm x 250 mm, 5 μm) chromatographic column, with 0.1% H3 PO4 water solution and acetonitrile as the mobile phase for gradient elution. The detection wavelength was 280 nm. HPLC fingerprint of Jinzhen oral solution was established to identify 17 common peaks in Jinzhen oral solution. The similarity of fingerprints of 10 batches of finished products was more than 0. 90. The established HPLC fingerprint has a better precision, reproducibility and stability, and can be applied in quality control of Jinzhen oral solution.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control

Triple negative breast cancer (TNBC), as a special molecular subtype of breast cancer, is non-responsive to endocrine therapy or commercially available targeted therapy. It is characterized by early recurrence, rapid progression and poor prognosis. This systemic and comprehensive overview was focused on recent progress on molecular subtyping of triple negative breast cancer and its possible clinical value, chemotherapeutic agents and chemotherapy regimens, and combination of chemotherapy with potential molecular targeting agents.

Objective To evaluate the Chinese version of Benefit Finding Scale (BFS) for breast cancer patients.Methods After consent of the author,according to the flow path of scales,we established the psychometric properties of the Chinese version of BUS,a convenience sample of 293 patients were recruited for evaluation.Results The Chinese version of BFS was comprised of 19 items,Cronbach's α coefficient was 0.911,test-retest reliability was 0.812,I-CVI was 0.833～1.000,S-CVI was 0.955.Meanwhile,exploratory analysis showed that the most interpretable solution consisted of 3 factors,the accumulative variance contribution which explained 55.101％ of variance of the total scale.The related validity showed that the correlative coefficient of Chinese version of BUS and Posttraumatic Growth Inventory (PTGI) was 0.745.Conclusions It suggests that the 19-item Chinese version of the BFS has good reliability and validity,which could be used as a research tool for measuring breast cancer patients' benefit finding.

Objective To investigate the molecular mechanism of Loureirin A mediated anti-hepatic fibrosis by evaluting its effects on proliferation , secretion ofα-smooth muscle actin (α-SMA) and transforming growth factor-beta1 (TGF-β1), and expression of rat hepatic stellate cells in vitro . Methods Primary hepatic stellate cells were isolated and cultured from Sprague-Dawley rats. After activating and inducing primary hepatic stellate cells from qHSC to aHSC, the activated hepatic stellate cells model in vitro was established. Then we observed the morphological changes of static hepatic stellate cells and activated hepatic stellate cells with inverted phase contrast microscope. Cultured hepatic stellate cells were treated with different concentrations of loureirin A and the inhibitory rate of HSCs proliferation was measured by MTT assay. The expression of Frizzled-4 was measured by western blot analysis. The content ofα-SMA and TGF-β1 in the cultured HSCs'supernatant were measured by enzyme-linked immunosorbent assay (ELISA) . Results Loureirin A the proliferation of inhibited activated hepatic stellate cells in a time-dose-dependent manner compared with the control group,IC50=0.30 μg/μL. After loureirinA treatment of the HSCs, western blot analysis showed that Frizzled-4 expression level was obviously lower than control group. Loureirin A also inhibitedα-SMA and TGFβ1 (P<0.05) secretion in the cultured HSCs'supernatant in different degree by the assay of ELISA. Conclusions The molecular mechanism of Loureirin A and Wnt signaling pathway mediated anti-hepatic fibrosis and anti-angiogenesis may involve down-regulation the expression of Frizzled-4, inhibiting the synthesis and secretion ofα-SMA,TGF-β1and the proliferation of HSCs.

rAdinbitor was cloned from Gloydius blomhoffi brevicaudus in the laboratory. Previous researches had proved that rAdinbitor could inhibit proliferation of C6 glioma cells as well as promote their apoptosis. The molecular mechanism of rAdinbitor’s effects on C6 cells need to be further studied. rAdinbitor was expressed in E. coli BL21/pET23b-adinbitor and purified with Ni Sepharose 6 Fast Flow. The purified protein was confirmed by Western blotting. C6 cells were induced with fibronectin (FN). The effects of rAdinbitor with different concentrations on the expression of FAK, MEK1/2 and Caspase-3 as well as on activity of FAK and ERK1/2 in FN-induced C6 cells were studied by immunoblotting and immunoprecipitation. Results showed that rAdinbitor with different concentrations could obviously reduce the expression of FAK and MEK1/2, increase the expression of Caspase-3, as well as decrease ERK1/2 phosphorylation; besides 10 mg/L rAdinbitor, other concentrations’ rAdinbitor could inhibit FAK phosphorylation obviously. All those effects were dose-dependent. Results indicate that the effects of rAdinbitor on decreasing expression and activity of FAK and inhibiting Ras-MAPK signaling pathway play an important role in suppressing the proliferation of C6. Furthermore, the increase in Caspase-3 expression implies that the increase in apoptosis of C6 cells might be due to the suppression of rAdinbitor on the activity of ILK and PI-3K/Akt pathway.

Objective To analyze the characteristics of breast medullary carcinoma in CEUS and to compare with pathologic features.Methods Morphologic characteristics of 13 breast medullary carcinomas in CEUS were analyzed.The diameter of mass before and after CEUS were compared.Parameters from time-intensity curves of masses were analyzed in contrast with peripheral breast parenchyma.All the results from CEUS analysis were compared with pathological manifestations.Results Breast medullary carcinoma was characterized as irregular shape (n=10),clear margins (n=11) and uniform enhancement (n=11) in CEUS.These characteristics were in accordance with their morphologic characters in pathology.The diameter of mass before and after CEUS had no significant defference (P=0.61),which was in accordance with expansive growth in pathology.In contrast with peripheral breast parenchyma,the arrival time and time to peak of breast medullary carcinoma were significantly shorter (P=0.034,0.021),and peak enhancement intensity was significantly stronger (P=0.005),which were in accordance with the increased vascular density and their uniform distribution,big arteries at the margin of masses in pathology.Conclusion Breast medullary carcinoma has distinguished characteristics in CEUS,which are in accordance with characters in pathology,and can be used as the basis in clinical diagnosis and differential diagnosis of breast medullary carcinoma.

Objective To study face recognition rule of Chinese subjects during judging the differences of facial recognition of race,gender and their joint property.Methods Eye tracking technique and moving window experimental method were used.Results The present results showed that,in facial race recognition,gender recognition and joint property recognition,the key area of face recognition was eyebrow and eye.In face recognition process,the dominant area in face of information acquired was right facial area.The key brain area of facial cognition was right brain area.Conclusion The recognition performance of face racial recognition is the best than face gender recognition and joint property recognition.Asymmetrical perception effect in human face area is appeared in facial recognition.

·AIM: To study the tear function changes in patients with pterygium before and after pterygium transposition.·METHODS: Twenty eyes of 20 patients were enrolled in this study.Schirmer I test and tear break-up time (BUT) were evaluated in patients before and after pterygium transposition.·RESULTS: There was no significant difference in the results of Schirmer I test before and after the surgery.The results of BUT were not significantly different before and up to 4 weeks after surgery. However, BUT prolonged significantly 6 weeks after surgery (P<0.05).·CONCLUSION: Pterygium transposition can improve the tear film function in patients with pterygium.

Background Antigen retrieval method is the key of improving the successful rate of immunohistochemical assay in paraffin sections.To study an available method of antigen retrieval is a goal to achieve both good immunochemistry result and preserving retinal proteins.ObjectiveThe aim of present study is to investigate tyrosin digestion,high-temperature heat pressure,water bath heating and microwave repair in antigen retrieval for retinal tissues.MethodsRetinal tissue was isolated and obtained from clean Chinchilla rabbits.Four hundreds retinal paraffin sections were prepared.Four kinds of antigen retrieval methods for retinal tissue including tyrosin digestion,high-temperature heat pressure,water bath heating and microwave repair were used respectively.The depigmentated retinal paraffin section without antigen retrieval was used as control.The positive rates of expression of CRALBP,Rhodopsin and opsin proteins were evaluated and compared among four kinds of antigen retrieval methods by immunochemistry.ResultsCRALBP,Rhodopsin and opsin protein was positively expressed in cytoplasm of retinal pigment epithelial cells and the outer segment of photoreceptor respectively.No significant difference was found in the positive expression rates of these three proteins among four kinds of antigen retrieval methods (P＞0.05),but the differences in tissue integrity and background staining were statistically significant (P＜0.01).The structural damage of retina included loss and pucker of scalera,crack of nucleus and abnormal background stain in high-temperature heat pressure method,water bath heating method and microwave retrieval method.However,stable CRALBP,Rhodopsin and opsin protein expression and strain effectiveness,clear background without unspecific staining and integrated tissue were seen in tyrosin digestion method.ConclusionDuring the clinical pathology analysis of retinal tissue,the application of tyrosin digestion in antigen retrieval could obtain a better effectiveness.