Study on Heavy Metal Detection in Soil with Improved EDXRF.

Due to the low precision and accuracy of trace heavy metals detection wiith traditional energy dispersive X-ray fluorescence (EDXRF) system, an improved EDXRF system is proposed. In order to reduce the influence of reflected rays, the sample is irradiated with the incident X-rays vertically, and the detector is placed in parallel with the sample's section. The sample is connected with detector through collimator. With improved EDXRF measures certified reference materials, the results show that the detection limit of the improved EDXRF system for Mo，Zn，Cu，Pb，Zr，Nb is 0.4，6.68，1.97，6.84，1.60，7.59 mg·kg-1 respectively and the logarithm deviation of each element in the standard samples is between 0 and 0.05. The RSD%(GBW) is less than 7 as the element content is more than three times of the detection limit, and it is below 15 when the element content is less than three times of the detection limit. The soil samples collected from Da Xing'an Ling region are applied to verify the improved EDXRF system. The proposed EDXRF system can improve the measurement accuracy of trace heavy metal detection in soil, satisfying the requirements of geologic exploration.

A meta-analysis and of clinical values of 11 blood biomarkers, such as AFP, DCP, and GP73 for diagnosis of hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma lacks ideal diagnostic biomarkers. There is a lack of scientific evaluation of relevant promising biomarkers as well. Therefore this study reanalyzes the related studies of 11 blood biomarkers of HCC, and compares the diagnostic value of these biomarkers for HCC systematically. METHODS: The relevant literatures on the diagnostic value in HCC of 11 blood indexes in recent 5 years were searched in PubMed, Embase, and Cochrane libraries. Data were extracted and analyzed. RESULTS: Finally, 83 literature studies were brought into meta-analysis. The pooled sensitivity and specificity of AFP were 0.61 and 0.87, respectively. The AUC of AFP were 0.78. The AUC and sum of sensitivity and specificity of the combination of AFP and other biomarkers were all significantly higher than that of AFP, including AFP + AFP-L3 + DCP, AFP + DCP, AFP/DCP, AFP + GPC3. Among other biomarkers, the AUC and sum of sensitivity and specificity of biomarkers including DCP, GPC3, GP73, Hsp90alpha, midkine, and OPN were significantly higher than that of AFP. In this study, GP73 had the highest sum of sensitivity and specificity (1.78) and AUC (0.95). CONCLUSIONS: The pooled sensitivity and specificity of AFP were 0.61 and 0.87, respectively. The AUC of AFP were 0.78. The combination of AFP and other biomarkers improved the diagnostic efficiency. The diagnostic value of biomarkers including DCP, GPC3, GP73, Hsp90alpha, midkine, and OPN was higher than that of AFP. GP73 had the best diagnostic value for HCC with the highest sum of sensitivity and specificity (1.78) and AUC (0.95).KEY MESSAGESThe pooled sensitivity and specificity of AFP were 0.61 and 0.87, respectively. The AUC of AFP were 0.78. The combination of AFP and other biomarkers improved the diagnostic efficiency of HCC.The diagnostic value of biomarkers including DCP, GPC3, GP73, Hsp90alpha, midkine, and OPN was higher than that of AFP.GP73 had the best diagnostic value for HCC.

[Effect of MXRA7 on the Biological Functions of Acute B Lymphoblastic Leukemia Cell Line REH].

OBJECTIVE: To discover the relationship between matrix remodeling associated 7 (MXRA7) and acute B lymphoblastic leukemia (B-ALL), and explore the effect of MXRA7 on the biological functions of B-ALL cell line REH. METHODS: The expression of MXRA7 in blood diseases was searched and analyzed through BloodSpot database. Real-time qPCR was used to detect the expression level of MXRA7 in B-ALL cell line 697 and REH cells. Lentivirus-mediated shRNA interference technology was utilized to knock down the expression of MXRA7 in REH cells. The effects of MXRA7 on the biological functions of REH cells were studied by in vitro experiments. Cell proliferation was detected by CCK-8 assay, cell cycle was detected by PI staining, cell apoptosis was detected by Annexin V and 7-AAD staining, and the expression of apoptosis pathway related proteins was detected by Western blot. RESULTS: Database analysis showed that MXRA7 was highly expressed in B-ALL patients, and real-time qPCR results showed that MXRA7 was also highly expressed in cell lines 697 and REH cells. Knockdown of MXRA7 in REH cells inhibited the cell proliferation and increased the percentage of G0/G1 phase cells. After treatment with cytarabine, the apoptotic ratio was increased in MXRA7-impaired REH cells, and the activation of caspase-3 and caspase-9 were also increased. CONCLUSION: Knockdown of MXRA7 can reduce the malignancy of REH cells by inhibiting the cell proliferation and increasing the sensitivity of REH cells to cytarabine. These results indicate MXRA7 may be as a novel target for the treatment of B-ALL, and the potential usefulness of MXRA7 in B-ALL deserves further investigation.

[Platforms are needed for innovative basic research in ophthalmology].

Basic research poses the cornerstone of technical innovation in all lines including medical sciences. Currently, there are shortages of professional scientists as well as technical supporting teams and facilities in the field of basic research of ophthalmology and visual science in China. Evaluation system and personnel policies are not supportive for innovative but high-risk-of-failure research projects. Discussion of reasons and possible solutions are given here to address these problems, aiming at promoting buildup of platforms hosting novel and important basic research in eye science in this country.

[The Effect of Matrix Remodeling Associated 7 (MXRA7) Expression on the Biological Function of SHI-1 Cells].

OBJECTIVE: To express matrix remodeling associated 7 (MXRA7) in the human acute myeloid leukemia SHI-1 cell line and to assess the role of MXRA7 in the biological function of SHI-1 cells. METHODS: The full-length cDNA sequence of human MXRA7 was synthesized and subcloned into the lentivirus shuttle vector pRRL-Venus. SHI-1 cells were transfected with the lentivirus which was packaged with 293T cells. The YFP-positive cells were sorted by flow cytometry and the stable cell lines were obtained by expanded culture. The expression and distribution of MXRA7 in SHI-1 cells were verified by real-time qPCR, Western blot and laser confocal techniques. Cell proliferation and cell cycle were measured by flow cytometry, and apoptosis was determined by Annexin V and 7-AAD staining. The expression of apoptosis related proteins were detected by Western blot. RESULTS: The stable SHI-1 cell line overexpressing MXRA7 was established successfully. Laser confocal analysis confirmed that MXRA7 was expressed in the cytoplasm of SHI-1 cells. Compared with the control cell line, the overexpression of MXRA7 showed no effect on the cell proliferation and cell cycle, but reduced the percentage of apoptosis cells induced by methotrexate. Moreover, the expression of BCL-2 protein was increased by overexpression of MXRA7, which can inhibit cell apoptosis. CONCLUSION: The SHI-1 stable cell line overexpressing MXRA7 was established successfully, and MXRA7 could inhibit drug-induced apoptosis through increasing the expression of BCL-2 protein.

[The preliminary study of establishing animal model of chronic allograft dysfunction after penetrating keratoplasty].

OBJECTIVE: To establish a murine model of chronic corneal allograft dysfunction (CCAD) after penetrating keratoplasty (PK). METHODS: Experimental study. PK model in mice: Semiallogeneic CB6F1 mice were obtained from matching of female BALB/c and male C57BL/6 mice. C57BL/6 (allogeneic group), CB6F1 (semiallogeneic group) and BALB/c (syngeneic group) grafts were transplanted orthotopically to BALB/c recipients respectively, and BALB/c mice as a control group. The follow-up time was more than 100 d, and graft survival time and corneal opacity score were monitored, and corneal endothelium were examined by alizarin red and PI/Hoechst stain. CD4(+) and CD8(+) T lymphocytes were examined by immunohistochemistry. Ultrastructure changes of the grafts were examined by electronmicroscopy. Log-rank test were used to compare survival curves. RESULTS: (1) Graft examination:Median graft survival times were 17.0 d, 85.5 d, > 100 d and > 100 d in allogeneic, semiallogeneic, syngeneic and control groups, respectively (F = 344.0, P < 0.01). (2) Immunohistochemistry examination: There were large amount of CD4(+) and CD8(+) T lymphocyte infiltration in allografts in allogeneic group at 3 weeks after PK; Few CD4(+) and CD8(+) T lymphocytes were observed in semiallogeneic group and syngeneic groups at 3 weeks after PK; CD4(+) and CD8(+) T lymphocyte infiltration was not observed in the control group. (3) Endothelium examination: The endothelium can not be counted because the blurred image after the alizarin red combined PI/Hoechst stain and apoptotic and necrotic cells can be seen in allogeneic group; the endothelial cell density decreased and few apoptosis can be detected in semiallogeneic and syngeneic groups; no apoptotic and necrotic endothelial cells were found in the control group. (4) Ultrastructural characteristic changes mainly include fibrosis formation and endothelium atrophy and degeneration in failed grafts in all transplanted groups by electron microscopy examination. Inflammation cells can only be found in the allogeneic group. CONCLUSIONS: Semiallogeneic and syngeneic transplantation groups present the changes similar to CCAD in clinical study, and both can be regarded as the model that permits molecular evaluation of CCAD.

Enhancement of tumor cell death by combining cisplatin with an oncolytic adenovirus carrying MDA-7/IL-24.

AIM: The aim of this study was to creatively implement a novel chemo-gene-virotherapeutic strategy and further strengthen the antitumor effect in cancer cells by the combined use of ZD55-IL-24 and cisplatin. METHODS: ZD55-IL-24 is an oncolytic adenovirus that harbors interleukin 24 (IL-24), which has a strong antitumor effect and was identified and evaluated by PCR, RT-PCR, and Western blot analysis. Enhancement of cancer cell death using a combination of ZD55-IL-24 and cisplatin was assessed in several cancer cell lines by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and cytopathic effect (CPE) assay. Apoptosis induction by treatment with ZD55-IL-24 and/or cisplatin was detected in BEL7404 and SMMC7721 by morphological evaluation, apoptotic cell staining, and flow cytometry analysis. In addition, negative effects on normal cells were evaluated in the L-02 cell line using the MTT assay, the CPE assay, morphological evaluation, apoptotic cell staining, and flow cytometry analysis. RESULTS: The combination of ZD55-IL-24 and cisplatin, which is superior to ZD55-IL-24, cisplatin, and ZD55-EGFP, as well as ZD55-EGFP plus cisplatin, resulted in a significantly increased effect. Most importantly, conjugation of ZD55-IL-24 with cisplatin had toxic effects equal to that of cisplatin and did not have overlapping toxicities in normal cells. CONCLUSION: This study showed that ZD55-IL-24 conjugated with cisplatin exhibited a remarkably increased cytotoxic and apoptosis-inducing effect in cancer cells and significantly reduced the toxicity in normal cells through the use of a reduced dose.

[Correlations of telomere length changing and pathogeny of keratoconus].

OBJECTIVE: To study telomere length, senescence-associated-beta-galactosidase (SA-beta-galactosidase) and senescence marker protein-30 (SMP-30) in the stromal cells of keratoconus or normal corneas respectively, aiming finding the association of these indexes with the phenotype of keratoconus. METHODS: Experiment research. 37 keratoconus lesions corneas were removed from 32 keratoconus patients who were operated in Shangdong Eye Institute between January 2006 and December 2006, and 20 normal corneas were collected from eye bank. The keratoconus corneas ages were from 13 to 34 years [mean ages (19 + or - 5) years] and the control group consists of 20 normal corneas donor ages from 9 to 30 years [mean ages (19 + or - 4) years]. And there was no statistical difference of ages between keratoconus and normal corneas. Southern blot method was utilized to detect telomere length of genomic DNA. SA-beta-galactosidase was detected by 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) staining method respectively in keratoconus and normal corneas. Isolated mRNA from keratoconus and normal corneas were reverse-transcribed to cDNA and SMP-30 was detected using PCR with specific primers (sense: 5' ccg tgg atg cct ttg act at 3'; anti-sense: 5' caa ctt cat gc tgct ttg ga 3'). To compare normal corneas and keratoconus corneas by histopathological study. Statistical analysis by t test. RESULTS: The telomere length in stromal cells in keratoconus corneas were from 10.29 to 14.12 kb, mean (11.54 + or - 1.41) kb, while that of normal corneas were from 12.64 to 15.32 kb, mean (13.45 + or - 0.99) kb. The difference of telomere length in stromal cells of keratoconus and normal corneas reached a statistical significant level (t = 4.753, P < 0.05). That means the telomere length of keratoconus stroma was shorter than that of normal corneal stroma. Light microscopy revealed that collagen fibers in keratoconus corneal stroma were arranged in an irregular manner. Cells density in keratoconus stroma appeared lower than in normal ones but the decrease was not significant. The staining of SA-beta-galactosidase in the keratoconus section was evident, but there was no staining in the normal corneas. SMP-30 was not detectable with RT-PCR method in either keratoconus or normal corneas. CONCLUSION: Telomeres in the keratoconus stromas manifest higher SA-beta-galactosidase than control, implying that improper senescence might be involved in pathogenesis of keratoconus.

An R124C mutation in TGFBI caused lattice corneal dystrophy type I with a variable phenotype in three Chinese families.

PURPOSE: A genetic and clinical study of three unrelated Chinese pedigrees with a variable phenotype of lattice corneal dystrophy type I (LCD I). METHODS: The eyes of the patients were examined by slit lamp microscopy, and other clinical records were also collected. Genomic DNA was extracted from peripheral leukocytes of the affected patients and their family members. Exons of the transforming growth factor beta induced TGFBI gene were amplified by polymerase chain reaction and directly sequenced to verify the mutation. Fifty healthy volunteers were analyzed as normal controls. RESULTS: Variable atypical clinical features of LCD I were found by slit lamp microscopy in these three Chinese pedigrees. A heterozygous single base-pair transition from C to T (C417T), leading to amino acid substitution (R124C) in the encoded TGFBI protein, was detected in all of the affected patients. No mutation was found in unaffected family members and 50 normal controls. CONCLUSIONS: Clinical features of Chinese patients with the same R124C mutation are quite variable even within the same family. Molecular genetic analysis of TGFBI can offer a rapid, accurate diagnosis of patients with atypical corneal dystrophies.

Theory of mind disability in major depression with or without psychotic symptoms: a componential view.

Previous reports have conceptualized theory of mind (ToM) as comprising two components and questioned whether ToM deficits are associated with psychotic symptoms. We investigated 33 nonpsychotic depressed inpatients, 23 psychotic depressed inpatients, and 53 normal controls with the following measures: Eyes Task, Faux pas Task, Verbal Fluency Test (VFT), Digit Span Test (DST) and WAIS-IQ. The depressed patients were also evaluated with the Beck Depression Inventory-II (BDI-II) and the Brief Psychiatric Rating Scale (BPRS). The nonpsychotic depressed patients and the psychotic depressed individuals were significantly impaired on tasks involving ToM social-perceptual and social-cognitive components, as well as the VFT. The psychotic depressed patients performed significantly worse than nonpsychotic depressed patients on ToM tasks. An association was found between ToM performances and both BPRS total and hostile-suspiciousness scores in the depressed group. Both of the ToM components were impaired in depressed patients. Similar mechanisms and neurobiological substrate may contribute to schizophrenia and major depression.

[Prevalence and predictors of acute stress disorder after earthquake: findings from Wenchuan earthquake in China].

OBJECTIVE: To investigate the prevalence and predictors of acute stress disorder (ASD) in the victims affected by Wenchuan earthquake in China. METHODS: A random clustered sampling method was used. Of 891 victims enrolled in the study, 874 were completely assessed with the ASD constructive questionnaire and diagnosed with DSM-IV criteria. Sociodemographic variables were obtained. Also, the major symptoms of ASD (i.e., general symptoms to a traumatic event; dissociative symptoms; re-experiencing symptoms; hyper-arousal symptoms; avoidance symptoms) were recorded. RESULTS: The incidence rate of ASD was 12.59% (110/874). The incidence rates of ASD for female and male were 15.16% (72/475) and 9.52% (38/399) respectively. There was a significant difference between female and male on the incidence rate of ASD (chi(2) = 6.26, P = 0.01). Logistic regression indicated that the ASD diagnosis was predicted by gender (beta = 0.58, P = 0.01, OR = 1.79), the condition of casualties of family members (beta = 0.60, P = 0.01, OR = 1.82), and the condition of sharp properties loss (beta = 1.02, P = 0.01, OR = 2.76). CONCLUSION: The major earthquake should have great influence on mental health of victims. The efforts to reduce casualties and property loss might help to prevent ASD. Further research is needed on gender difference among traumatic events.

[Effect of Tongdatang Serial Recipe on antipsychotic drug-induced galactorrhea-amenorrhea syndrome].

OBJECTIVE: To observe the clinical effect of self-formulated Tongdatang serial recipe (TDT) in treating antipsychotic drug-induced galactorrhea-amenorrhea syndrome (GAS). METHODS: One hundred female schizophrenic patients with antipsychotic drug-induced GAS were selected and equally assigned to the treatment group and the control group at random. Both received antipsychotic drug-therapy, but combined with TDT and placebo respectively. The efficacy was evaluated by determining prolactin level before, 4 and 8 weeks after treatment. RESULTS: The treatment course was completed in 96% of patients. Therapeutic efficacy on the 49 patients of the treatment group was cured in 31 (63.3%), markedly effective in 11 (22.4%), effective in 4 (8.2%) and ineffective in 3 (6.1%), with total effective rate of 93.9%, while in 47 patients of the control group, the corresponding cases (%) was 0, 3 (6.4%) , 7 (14.9%) and 37 (78.7%), respectively, with the total effective rate of 21.3%. Prolactin levels in the two groups were insignificantly different before treatment, it reduced in the treatment group after treatment (P < 0.01), and the decrement in the treatment group was more significant than that in the control group (P < 0.05). CONCLUSION: Satisfactory effect could achieved by using TDT for treatment of antipsychotic drug-induced GAS.

Family history of suicide and high motor impulsivity distinguish suicide attempters from suicide ideators among college students.

Suicide in college students has become an important public health issue in China. The aim of this study was to identify the differences between suicide attempters and suicide ideators based on a cross-sectional survey. Our results indicate that although female gender, positive screening for psychiatric illness, positive family history of suicide, elevated overall impulsivity, and elevated motor impulsivity were correlated with suicidal ideation, only positive family history of suicide and high motor impulsivity could differentiate suicide attempters from suicidal ideators. Future research with a longitudinal and prospective study design should be conducted to confirm these findings.

The involvement of proline-rich protein Mus musculus predicted gene 4736 in ocular surface functions.

AIM: To research the two homologous predicted proline-rich protein genes, Mus musculus predicted gene 4736 (MP4) and proline-rich protein BstNI subfamily 1 (Prb1) which were significantly upregulated in cultured corneal organs when encountering fungal pathogen preparations. This study was to confirm the expression and potential functions of these two genes in ocular surface. METHODS: A Pseudomonas aeruginosa keratitis model was established in Balb/c mice. One day post infection, mRNA level of MP4 was measured using real-time polymerase chain reaction (PCR), and MP4 protein detected by immunohistochemistry (IHC) or Western blot using a customized polyclonal anti-MP4 antibody preparation. Lacrimal glands from normal mice were also subjected to IHC staining for MP4. An online bioinformatics program, BioGPS, was utilized to screen public data to determine other potential locations of MP4. RESULTS: One day after keratitis induction, MP4 was upregulated in the corneas at both mRNA level as measured using real-time PCR and protein levels as measured using Western blot and IHC. BioGPS analysis of public data suggested that the MP4 gene was most abundantly expressed in the lacrimal glands, and IHC revealed that normal murine lacrimal glands were positive for MP4 staining. CONCLUSION: MP4 and Prb1 are closely related with the physiology and pathological processes of the ocular surface. Considering the significance of ocular surface abnormalities like dry eye, we propose that MP4 and Prb1 contribute to homeostasis of ocular surface, and deserve more extensive functional and disease correlation studies.

[Platelets Decrease the Sensitivity of Leukemia Cell L1210 to Multiple Drugs via Activiting The AKT and ERK Signalling Pathway].

OBJECTIVE: To explore the effect of platelets on signal transducers and the sensitivity of leukemia cells to chemotherapeuticl drugs in leukemia cells L1210. METHODS: Murine platelets were prepared and cocultured with leukemia L1210 cells, and the aggregation between them was observed by flow cytometry. The levels of several transducer proteins in leukemia cells were analyzed with Western blot. In some experiments, methotrexate, vincristine or doxorubicin was added to the coculture system and the cell proliferation was measured by using CCK8 colorigenic methods to detect the sensitivity of leukemia cells to the therapeuticals drugs. RESULTS: Platelets, either freshly prepared or fixed with 1% paraformaldehyde, aggregated with leukemia cells. Both fresh and fixed platelets increased the level of phosphorylation of AKT and ERK in leukemia cells as measured with Western blot. Also, platelets significantly decreased the sensitivity of 3 therapeutics to L1210 cells. CONCLUSION: Platelets may bind with L1210 cells and decrease the sensitivity of the leukemia cells to chemotherapeutics, possibly by activating the AKT and ERK signaling pathways.

[Progress in the study of epidemiology and etiology of age-related macular degeneration].

Age-related macular degeneration is one of the leading causes of blindness in elder population, and many studies have been done on its epidemiology, etiology and pathogenesis. However, many issues are still open to question due to the complexity of the disease development and lack of the adequate animal model.

Herpes simplex virus-1 infection or Simian virus 40-mediated immortalization of corneal cells causes permanent translocation of NLRP3 to the nuclei.

AIM: To investigate into the potential involvement of pyrin containing 3 gene (NLRP3), a member of the nucleotide-binding oligomerization domain-like receptors with cytosolic pattern recognition, in the host defense of corneas against viruses. METHODS: The herpes viral keratitis model was utilized in BALB/c mice with inoculation of herpes simplex virus-1 (HSV-1). Corneal tissues removed during therapy of patients with viral keratitis as well as a Simian vacuolating virus 40 (SV40)-immortalized human corneal epithelial cell line were also examined. Immunohistochemistry was used to detect NLRP3 in these subjects, focusing on their distribution in tissue or cells. Western blot was used to measure the level of NLRP3 and another two related molecules in NLPR3 inflammasome, namely caspase-1 and IL-1ß. RESULTS: The NLRP3 activation induced by HSV-1 infection in corneas was accompanied with redistribution of NLRP3 from the cytoplasm to the nucleus in both murine and human corneal epithelial cells. Furthermore, in the SV40-immortalized human corneal epithelial cells, NLRP3 was exclusively located in the nucleus, and treatment of the cells with high concentration of extracellular potassium (known as an inhibitor of NLRP3 activation) effectively drove NLRP3 back to the cytoplasm as reflected by both immunohistochemistry and Western blot. CONCLUSION: It is proposed that herpes virus infection activates and causes redistribution of NLRP3 to nuclei. Whether this NLRP3 translocation occurs with other viral infections and in other cell types merit further study.

[Expression Changes of Matrix Reconstitution-related Genes in Bone Marrow during 5-Fluorouracil-Induced Hemopoietic Injury and Their Significance].

OBJECTIVE: To explore the expression change of the genes related with matrix reconstitution during the injury and reconstitution of murine bone marrow following treatment with 5-fluorouracil (5-FU). METHODS: C57BL/6 mice received intraperitoneal injection of 5-FU (200 mg/kg), and peripheral blood cell counts were monitored at 3, 6, 9, 15, 21, 27 days after treatment. Bone marrow cells were harvested at these times for total RNA extraction using TRIzol. Reverse transcriptions in combination with real-time PCR were performed for detecting expression of genes related with matrix reconstitution, including ECM-1, MMP-2, MMP-3, MMP-13 and TIMP-1. RESULTS: After injection of 5-FU, the numbers of three line cells in peripheral blood (i.e. RBC, WBC and platelets) decreased and then recovered with differential dynamics. Similarly, RT-qPCR revealed that all the 5 detected gene expressions were significantly up-regulated during the injury. The mRNA expression of MMP-2 reached to peak at day 3 while the other genes reached to peak at day 6. MMP-3 has a low expression when compared with others, but its expression increased significantly after injury. CONCLUSION: In 5-FU induced hematopoietic injury and reconstitution model, matrix reconstitution-related genes may play an important role in hematopoietic reconstitution, but different genes play different roles at various time, and cooperate with each other for hematopoietic reconstitution.

Serum amyloid A and pairing formyl peptide receptor 2 are expressed in corneas and involved in inflammation-mediated neovascularization.

AIM: To solidify the involvement of Saa-related pathway in corneal neovascularization (CorNV). The pathogenesis of inflammatory CorNV is not fully understood yet, and our previous study implicated that serum amyloid A (Saa) 1 (Saa1) and Saa3 were among the genes up-regulated upon CorNV induction in mice. METHODS: Microarray data obtained during our profiling project on CorNV were analyzed for the genes encoding the four SAA family members (Saa1-4), six reported SAA receptors (formyl peptide receptor 2, Tlr2, Tlr4, Cd36, Scarb1, P2rx7) and seven matrix metallopeptidases (Mmp) 1a, 1b, 2, 3, 9, 10, 13 reportedly to be expressed upon SAA pathway activation. The baseline expression or changes of interested genes were further confirmed in animals with CorNV using molecular or histological methods. CorNV was induced in Balb/c and C57BL/6 mice by placing either three interrupted 10-0 sutures or a 2 mm filter paper soaked with sodium hydroxide in the central area of the cornea. At desired time points, the corneas were harvested for histology examination or for extraction of mRNA and protein. The mRNA levels of Saa1, Saa3, Fpr2, Mmp2 and Mmp3 in corneas were detected using quantitative reverse transcription-PCR, and SAA3 protein in tissues detected using immunohistochemistry or western blotting. RESULTS: Microarray data analysis revealed that Saa1, Saa3, Fpr2, Mmp2, Mmp3 messengers were readily detected in normal corneas and significantly up-regulated upon CorNV induction. The changes of these five genes were confirmed with real-time PCR assay. On the contrary, other SAA members (Saa2, Saa4), other SAA receptors (Tlr2, Tlr4, Cd36, P2rx7, etc), or other Mmps (Mmp1a, Mmp1b, Mmp9, Mmp10, Mmp13) did not show consistent changes. Immunohistochemistry study and western blotting further confirmed the expression of SAA3 products in normal corneas as well as their up-regulation in corneas with CorNV. CONCLUSION: SAA-FPR2 pathway composing genes were expressed in normal murine corneas and, upon inflammatory stimuli challenge to the corneas, their expressions were up-regulated, suggesting their roles in pathogenesis of CorNV. The potential usefulness of SAA-FPR2 targets in future management of CorNV-related diseases deserves investigation.

Comparison of the therapeutic effects of extracts from Spirulina platensis and amnion membrane on inflammation-associated corneal neovascularization.

AIM: To compare the therapeutic effects of polysaccharide extract from Spirulina platensis (PSP) and extract from amnion membrane (AME) on alkali burn-induced corneal neovascularization (CorNV). METHODS: PSP and AME were extracted from dry powder of Spirulina platensis and human aminion membrane respectively. Murine CorNV was induced by applying 1N sodiumhydroxide (NaOH) solution directly on the mice corneas. PSP and AME extracts were administered topically on the corneas 4 times daily for 7 days. The therapy effects of PSP and AME extracts were evaluated daily using slit-lamp. At the end of the therapy, corneas were harvested for H&E staining, masson trichrome staining, immunohistochemical study, and semi-quantification reverse transcriptive PCR (RT-PCR) was utilized for measurement of inflammation-related molecules. RESULTS: Topical application of PSP extract had significant therapeutic effects on CorNV that could be shown in various assays of the corneas. Compared with AME extract, PSP extract treatment was more effective in suppressing CorNV in terms of vessel length and levels of cluster of differentiation 31 (CD31) proteins or the angiogenesis related genes like vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP2) and matrix metalloproteinase-9 (MMP9). PSP also inhibited inflammation more markedly by more effectively inhibiting mononuclear and polymorphonuclear cells infiltration into the corneal stroma and reducing levels of stromal cell-derived factor-1 (SDF1), tumor necrosis factor-alpha (TNFα) and macrophage inflammatory protein-3 (MIP3a). In additon, corneas of PSP group had a more regular and compact architecture of collagen with thinner corneal thickness than in the AME group. CONCLUSION: Polysaccharide extract from Spirulina platensis inhibited alkali burn-induced inflammation and CorNV more effectively than AME extract at the studied doses, thus may be used for the therapy of corneal diseases involving neovascularization and inflammation.