The CD2-CD58 axis: A novel marker predicting poor prognosis in patients with low-grade gliomas and potential therapeutic approaches.

INTRODUCTION: Low-grade gliomas (LGGs) are currently considered a premalignant condition for high-grade gliomas (HGGs) and are characterized by a relatively intact immune system. Immunotherapeutic modalities may offer a safe and effective treatment option for these patients. However, the CD2-CD58 axis, an important component of the immunological synapse, remains unknown in LGG. METHODS: RNA-seq data from TCGA databases were analyzed. Immune cell infiltration was determined using a single-sample gene set enrichment analysis (ssGSEA) based on integrated immune gene sets from published studies. Kaplan-Meier survival analysis, univariate and multivariate logistic analysis, and the ESTIMATE algorithm were employed to evaluate the impact of the CD2-CD58 axis on adult LGG patients. RESULTS: The expression of the CD2-CD58 axis was found to be elevated with increasing of WHO grade (p < .05). Uni- and multi-variable logistic analysis demonstrated that age, WHO grade, and CD58 levels were associated with poor prognosis in LGG patients with (p < .01). MetaSape pathways analysis revealed the involvement of CD58 in regulating T cell activation, leukocyte-mediated immunity, and the positive regulation of cell activation in WHO grade II and III. CD58 expression correlated with infiltrations of CD4+ lymphocytes, NK cells, and macrophages cells. The ESTIMATE algorithm indicated that patients with high CD58 expression had significantly higher immune scores compared with low CD58 expression in WHO grade II/III, but no statistical difference was observed in WHO grade IV (p < .05). Furthermore, correlation analysis demonstrated the significant association between CD58 and CD274 (r = 0.581, p < .001), HAVCR2 (r = 0.58i7, p < .001), and LGALS9 (r = 0.566, p < .001). Immunohistochemical staining further confirmed the relationship of CD58, HAVCR2, WHO grade, and prognosis in grade II and III patients. CONCLUSION: Overall, our findings highlight the significant association between the CD2-CD58 axis and poor survival in LGG patients. High CD58 expression is implicated in T cell-mediated immune responses as an immunosuppressive factor and affect inhibitory immune checkpoint genes.

Fractionated carbon dioxide (CO2) laser treatment contributes to trans-nail penetration of rhodamine B and changes of cytokine microenvironment.

This study is to determine the role of the fractional CO2 laser in topical drug delivery and the impact of local immune responses. Experimental rabbit nails were treated with fractionated CO2 laser at varied fluencies of 20 mJ, 25 mJ, and 30 mJ and half of which were coated with rhodamine B (RhB). Histological examination was performed by hematoxylin and eosin staining; the penetration of RhB was assessed by the use of confocal laser scanning microscopy; and the expressions of IFN-γ and IL-4 mRNA in situ were detected by means of qPCR at 12 h, 24 h, 3 days, and 7 days post-laser irritation. The fractional CO2 laser could generate microscopic treatment zones in nail plates, and the depths of these micropores as well as the permeation of RhB in nails increased significantly in an energy-dependent manner. Importantly, the laser irritation led an upregulation of local IFN-γ mRNA expression accompanied by a downregulation of IL-4 mRNA expression. The ultrapulsed ablative fractionated CO2 laser may assist topical drug delivery, and may drive stronger local Th1 responses due to an imbalance of IFN-γ/IL-4 expressions, suggesting that the combination of ablative fractionated CO2 laser with topical agents would be an effective option for the treatment of onychomycosis.

Dysregulated miR-137 and its target EGFR contribute to the progression of pituitary adenomas.

Pituitary adenomas (PAs) hypersecrete hormones or cause mass effect symptoms, with 10%-35% patients showing resistance to standard therapies. Targeting epidermal growth factor receptor (EGFR) has significantly improved the clinical outcome in many cancers. In this study, immunochemistry results showed that EGFR associated H-scores in 116 PA samples were higher than those in pituitary glands, and that p21, p27-and Wif-1 associated H-scores were lower (P < 0.05 for all). Patients with high levels of EGFR had increased PA invasion, lower total resection, and lower p21 and p27 expression than those with low levels of EGFR expression. Dual-luciferase reporter gene assays showed that EGFR was the target gene of miR-137, and miR-137 mimic could inhibit the cell proliferation of GH3 cells and induce apoptosis and G1-phase arrest of GH3 cells. A combination of miR-137 mimic and AZD9291 had stronger inhibition on GH3 cells compared with miR-137 mimic or AZD9291 alone; furthermore, miR-137 inhibitor partially reversed the inhibition of AZD9291. p21 and p27 were shown to be involved in the miR-137- and AZD9291-mediated effects on GH3 cells. In all, activation of EGFR in PAs was related to tumor invasive behavior, which reduced the total resection of PAs in patients. A combination of miR-137 and AZD9291 provided a potential treatment for PAs, especially for patients who show resistance to standard treatment.

The absence of PRDM2 involved the tumorigenesis of somatotroph adenomas through regulating c-Myc.

Somatotroph adenoma is the main cause of acromegaly which have peripheral signs with growth of soft tissues and multiple comorbidities. Surgery and adjuvant therapy with somatostatin analogs (SSA) fail in more than 25% of patients. PRDM2, a tumor suppressor, plays an important role in cancer and obesity, including pituitary adenomas. In this study, we analyze the correlation of PRDM2 and oncogene c-Myc in 70 somatotroph adenomas according immunohistochemical staining, furthermore, we probed that whether PRDM2 participates in c-Myc signaling pathway in vitro experiment. 70 somatotroph adenomas patients were divided into low patients and high patients according to median of H-score of PRDM2 or c-Myc. Low PRDM2 patients had higher risk of invasive behavior, larger tumor volume and recurrence chance than high PRDM2 group (P = 0.015, P = 0.031, P = 0.017). High c-Myc patients had higher risk of invasive behavior, larger tumor volume and recurrence chance than low c-Myc group (P = 0.012, P = 0.002, P = 0.015). It was a negative correlation between H-score of PRDM2 and c-Myc (PRDM2 = -0.163 × c-Myc + 67.11, r = -0.407). The ability of cell proliferation was declined in a time dependent manner after overexpression of PRDM2 (PRDM2 group) compared to that in control GH3 cells (P < 0.05). Through flow cytometry assay, PRDM2 could induce the apoptosis and G2/M arrest in GH3 cell (both p < 0.05). Transwell experiment proved less trans-membrane cells in PRDM2 group than those in control group (415 ± 76 vs 145 ± 37, P < 0.01). RT-PCR and western blot both proved PRDM2 could inhibit the level c-Myc and elevate the levels of CDKN1A and CDKN1B. Combined with c-Myc inhibitor 10058-F4, PRDM2 further inhibited cell proliferation and induced more apoptosis in GH3 cell. Taken together, we found that PRDM2 negatively regulated the expression of c-Myc in somatotroph adenomas, and testified the synergism between PRDM2 gene therapy and c-Myc inhibitor in vitro experiment.

[Design of schistosomiasis surveillance sites based on Spatial Kluster Analysis by Tree Edge Removal (SKATER) method: an exploratory study].

OBJECTIVE: To develop a method for designing schistosomiasis surveillance sites, so as to improve the efficiency and quality of schitsosomiasis surveillance. METHODS: By using the minimum spanning tree-based Spatial Kluster Analysis by Tree Edge Removal (SKATER) method, spatially constrained clustering was performed upon 31 historical schistosomiasis-endemic counties (districts) in Anhui Province. A surveillance site was selected from each cluster to evaluate the representativeness and surveillance efficiency of these cluster-based surveillance sites for the endemic situation of schistosomiassi in Anhui Province, and to compare the surveillance efficiency with local national schistosomiasis surveillance sites. RESULTS: There was no significant difference in the environmental factors between the cluster-based schistosomiasis surveillance sites and the whole region, showing a high homogeneity. If the same number of schistosomiasis surveillance sites was selected, there was no significant difference between the cluster-based surveillance sites and national schistosomiasis surveillance sites in the efficiency of the mean risk and long-term trend of schistosomiasis surveillance in Anhui Province; however, the cluster-based surveillance sites were superior to the national schistosomiasis surveillance sites for the prediction and estimation of the endemic situation of schistosomiasis in the unmonitored areas. CONCLUSIONS: The SKATER-based selection of schistosomiasis surveillance sites may better represent the endemic situation of schistosomiasis in Anhui Province, which may serve as an effective supplement for the conventional method of selecting schistosomiasis surveillance sites.

[Spatial - temporal clustering analysis of schistosomiasis transmission risk in Yunnan Province from 2004 to 2015].

OBJECTIVE: To explore the spatial-temporal clustering distribution of schistosomiasis transmission risk in Yunnan Province from 2004 to 2015, so as to provide scientific evidence for the future schistosomiasis control and consolidation of the control achievements. METHODS: All data pertaining to endemic situation of human and bovine schistosomiasis and snail survey at a township level in Yunnan Province from 2004 to 2015 were collected. A descriptive epidemiological method and Joinpoint model were used to describe the changing trends of Schistosoma japonicum infections in humans, bovine and snails, and the hotspots and clusters of schistosomiasis transmission risk were identified using spatial autocorrelation analysis, hotspots analysis and retrospective space-time scan statistic in Yunnan Province from 2004 to 2015. RESULTS: The prevalence of S. japonicum infections appeared a continuous decline in humans, bovine and snails in Yunnan Province from 2004 to 2015, and the estimated number of schistosomiasis cases reduced from 43 056 in 2004 to 756 in 2015, with a decline rate of 98.24%. There were no acute cases since 2008 and no infected snails since 2014 in Yunnan Province. There were significant differences in the changing trends of human and bovine S. japonicum infections in Yunnan Province between 2012 and 2015 and between 2013 and 2015, respectively using the Joinpoint model (P < 0.05). In addition, there was a spatial autocorrelation in human S. japonicum infections in Yunnan Province from 2004 to 2013 (P < 0.01), and the hotspots areas for human S. japonicum infections were mainly distributed in some townships from Dali City, Weishan County and Eryuan County. Retrospective spatial-temporal scanning revealed that S. japonicum human, bovine and snail infections were clustered in 23, 15, 4 townships from Dali City, Weishan County, Eryuan County, Nanjian County and Heqing County, respectively, with relative risks of 6.25 to 28.75 (P < 0.01), which was almost consistent with the cluster areas detected by hotspots analysis. CONCLUSIONS: The endemic situation of schistosomiasis significantly reduced in Yunnan Province from 2004 to 2015; however, there is still a risk of schistosomiasis transmission. The monitoring and control of schistosomiasis should be intensified in the future in Yunnan Province.

[Observation on growth and reproduction of various Leishmania isolates in different culture media].

OBJECTIVE: To compare the growth and reproduction of the promastigotes of Leishmania isolates from various endemic areas of visceral leishmaniasis in China in various culture media, so as to provide experimental evidence for selecting an appropriate medium for the culture of Leishmania. METHODS: A total of 3 × 105 promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates were inoculated into 1 mL NNN medium, 1 mL M199 medium supplemented with 20% fetal bovine serum medium, 1 mL M199 medium supplemented with 20% horse serum medium, and 1 mL brain heart infusion medium containing heme, respectively. All media were placed at 22 ℃ under a sterile condition, and the number of promastigotes was counted continuously for 8 days under a microscope. The growth curve was plotted for the three Leishmania isolates. RESULTS: The promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates all grew and reproduced in the NNN medium, the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium. The number of promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates was all significantly higher in the NNN medium than in the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium at various time points of culture (all P values < 0.05), and the number of promastigotes of the KS-2 isolate was all significantly greater than that of the Cy and JIASHI-5 isolates in the NNN medium, the M199 medium supplemented with 20% fetal bovine serum medium, and the M199 medium supplemented with 20% horse serum medium at various time points of culture (all P values < 0.05). In ad dition, the promastigotes of KS-2, Cy and JIASHI-5 Leishmania isolates failed to grow and reproduce in the brain heart infusion medium. CONCLUSIONS: The growth and reproduction of the promastigotes of various Leishmania isolates from various endemic areas of visceral leishmaniasis in China vary in the same culture medium, and the growth and reproduction of a Leishmania isolate vary in different culture media. The NNN medium best fits for the culture of Leishmania isolates in the endemic areas of visceral leishmaniasis in China.

MiR-137's Tumor Suppression on Prolactinomas by Targeting MITF and Modulating Wnt Signaling Pathway.

CONTEXT: Prolactinomas are the most common functional pituitary adenomas; the aggressive tumors still present challenge to clinicians. Aberrant expression of miRNAs has been functionally associated with prolactinomas. OBJECTIVE: Here we explored the role of miR-137 on the proliferation, invasion, and apoptosis of prolactinomas and its possible mechanism. RESULTS: Low expression of miR-137 was correlated with the invasive behavior of human prolactinomas and predicted high recurrence. MiR-137 inhibited cell proliferation, invasion, and survivals of MMQ and GH3 cells and reduced tumor volume in F344 rat prolactinomas. The luciferase reporter assay confirmed that microphthalmia-associated transcription factor (MITF) was the direct target of miR-137. In addition, miR-137 mimics could inhibit MITF expression in vivo and in vitro. Upregulation of MITF expression promoted cell proliferation, invasion, and survivals and reversed the antitumor effect of miR-137 in vivo and in vitro. Furthermore, miR-137 could also upregulate wnt-inhibitory factor-1 and inhibit nuclear translocation of ß-catenin. Upregulation of wnt-inhibitory factor-1 with decitabine can enhance the inhibition on cell proliferation of miR-137. A glycogen synthase kinase-3 inhibitor, SB 216763, promoted cell proliferation by upregulation of total/cytoplasmic/nuclear ß-catenin and reversed tumor suppression of miR-137 mimics. CONCLUSIONS: Our data suggest that miR-137 possesses a tumor invasive suppressor function with a prognostic value in prolactinomas by targeting MITF and modulating Wnt-ß-catenin signaling pathway.

[Investigation of Oncomelania hupensis snails in Anhui Province in 2016].

OBJECTIVE: To investigate the status of Oncomelania hupensis snails in Anhui Province in 2016, so as to provide the evidence for formulating the 13th Five-Year Plans for Schistosomiasis Control. METHODS: In 2016, a snail survey was conducted in Anhui Province according to the National Programme of the Oncomelania Snail Survey, covering all snail habitats or historical snail habitats and suspected environments of snail breeding, and the results of the survey were analyzed. RESULTS: In 2016, 22 848 environments were surveyed, and no schistosome-infected snails were found. There were 17 927 historical snail habitats in whole province, among which, the environments without changes, changed partly and changed completely accounted for 71.2%, 19.3% and 9.5%, respectively. The area of historical snail habitats was 1.410 billion m2. There were 4 830 environments with snail habitats covering an area of 0.265 billion m2 in 38 counties of 7 cities, including newly emerging area of 1 287.65 hm2 and reemerging area of 1 375.32 hm2. The density of living snails was 0.392 0 snails/0.1 m2, and the rate of frame with living snails was 12.93%. The type of marshland and lake regions, and the type of hilly and mountainous regions accounted for 22.4% and 77.6% of number of snail habitats, and accounted for 86.7% and 13.3% of areas of snail habitats, respectively. Among the different types of vegetation in snail habitats, grass was superior owe to accounting for 82.2% of the number of snail habitats, 57.3% of the area with snail habitats, and the highest density of living snails (0.413 9 snails/0.1 m2). Among the different types of environments in snail habitats, the ditch was superior owe to accounting for 56.8% of the number of snail habitats, the highest density of living snails (0.570 3 snails/0.1m2) and the highest rate of frame with living snails (18.57%), and the beach was superior owe to accounting for 87.8% of the area with snail habitats. In Anhui Province, the first year of snails and schistosome-infected snails being found was 1950 and 1952, respectively, and the latest year of schistosome-infected snails being found was 2012. The map showed that the most environments with snail habitats were distributed along the Yangtze River in Anhui Province. CONCLUSIONS: For the first time, the investigation has built the database and map of snail spatial distribution, which truly reflect the historic and current status of snail distribution in Anhui Province and can provide the evidence for formulating the 13th Five-Year Plans for Schistosomiasis Control and improving the schistosomiasis prevention and control work in the future.

Detection of circulating tumor cells in patients with pituitary tumors.

BACKGROUND: Circulating tumor cells (CTCs) are tumor cells that have shed from a primary tumor and circulate in the peripheral blood. Recent experimental and clinical studies show that CTCs can be detected in early-stage disease. CASE PRESENTATION: We report three cases of pituitary adenoma (PA) in which tumor cells with particles were detected in the interstitial vascular compartment by transmission electron microscopy. Tumors were completely resected. Immunohistochemical analysis showed a ß-catenin score of 10.5 ± 1.5 in the three cases with CTCs compared with 2.4 ± 0.5 in 24 control adenomas. The Ki-67 labeling index was 2.1 ± 0.7 in CTCs vs. 0.2 ± 0.3 in control cases (p = 0.043), and the p53 score was 4.33 ± 1.3 vs. 0.31 ± 0.17 (p = 0.000). The E-cadherin score did not differ significantly between the two groups. CONCLUSIONS: CTCs can be detected in benign tumors such as PAs and not only in late-stage malignant tumors with apparent distant metastases. The present findings suggest that pituitary carcinomas develop from adenomas.

[Application of infrared camera technique in investigation of wild animals as infectious source of schistosomiasis].

OBJECTIVE: To investigate the species and activities of wild animals infected with schistosome in hilly and mountain areas by using an infrared camera technique, so as to provide the evidence for the surveillance of schistosomiasis in these areas. METHODS: Six infrared cameras were selected and placed in 6 environments of the risk monitoring points of schistosomiasis in Shitai County in Anhui Province. The species and activities of the wild animals in the 6 environments were observed through the photographs and videos taken by the cameras. RESULTS: Through 5 day's monitoring, 3 wild mammals, such as voles, hares and wild boars, were found in 4 monitoring environments, of which voles were found at 2 environments with snails, and hares, wild boars and voles were found in 2 environments adjacent to environments with snails respectively. The monitoring showed that the vole activity was most frequent in the monitored environment. CONCLUSIONS: The use of infrared camera technique has a good effect in the investigation of wild animal infectious source of schistosomiasis, and it is also suitable for the monitoring work in other types of environments.

[Comparative proteomics analysis of Leishmania infantum strains isolated from two epidemiological regions of visceral leishmaniasis in China].

OBJECTIVE: To analyze the protein abundance differences between two Leishmania infantum strains isolated from different epidemiological types of visceral leishmaniasis in China by comparative proteomics method. METHODS: Tryptic digests of total proteins were analyzed by using liquid chromatography-tandem mass spectrometry (LC-MS/MS), followed by label-free quantitative differential expression analysis. The MS data were analyzed with MaxQuant software (ver 1.3.0.5) against data base. RESULTS: This study resulted in the identification of 4 274 proteins across two strains (JIASHI-5 and SC6) . Of these, 1 219 differentially expressed proteins (ratio ＞ 2.0 or ＜ 0.5, P < 0.05) were identified. Considering the proteins differentially or uniquely expressed in the strains, 550 proteins were only found in the JIASHI-5 strain, and 174 proteins were only found in the SC6 strain. Totally 495 differentially proteins were expressed in the two groups, among which 328 proteins were down-regulated and 167 proteins were up-regulated in SC6 strain. Some of the identified differentially expressed proteins were demonstrated and they involved in energy metabolism, stress response, prolonging the lifetime of the infected host cell and survival and proliferation in virulent strains. CONCLUSIONS: This study reveals a group of differentially expressed proteins and the related biologic function that may lay the foundation for screening and identification of the key Leishmania molecules relative to pathogenicity.

Noninvasive evaluation of corneal viscoelasticity based on displacement in response to acoustic radiation force.

Noninvasive assessment of corneal mechanical properties in vivo will help to the understanding of the pathogenesis and early diagnosis of ectatic corneal disorders. This study presented a noninvasive method that assesses the corneal biomechanical properties by exciting the cornea with acoustic radiation force and monitoring its displacement using a dual-frequency confocal transducer. A 3.85-MHz pushing element was used to induce localized tissue displacement, and the displacement was detected by the 12.8-MHz detecting elements using pulse-echo methods. Under constant acoustic radiation force, 0the tissue displacement are directly correlated with tissue biomechanical properties, a set of parameters were extracted from local displacement waveform, including relaxation time constant (τ), relative elasticity (RE) and relative viscosity (RV). In order to obtain corneal samples with different mechanical properties, the fresh bovine eyes were performed by collagen cross-linking (CXL). The result indicated that the estimated τ in untreated corneas were statistically significantly different (p<;0.05) from those of treated corneas and the estimation of τ varied significantly with different degrees of CXL. It is possible to develop a non-invasive, effective and efficient method with high spatial resolution.

[Interpretation of Diagnostic Criteria for Kala-azar].

Kala-azar was once transmitted in the northern area of the Yangtze River in China, including 16 provinces (cities or autonomous regions). Through the great continuing prevention and control effort, this disease has been effectively controlled in the most of endemic areas. However, because the epidemic factors of the disease are complex, this disease still transmits or sporadically occurs in the western part of China, including 60 counties of Xinjiang, Gansu, Sichuan, Inner Mongolia, Shanxi, and Shaanxi provinces (autonomous regions). Following the Management Measures for Health Criteria, the Diagnostic Criteria for Kala-azar (WS 258-2006) was compiled by the ex-Ministry of Health of the People's Republic of China and it was issued in April 7, 2006 and implemented in December 1, 2006. The Criteria consists of six parts, including the application range, terms and definitions, diagnostic principle, diagnostic standard, and differential diagnosis. Two informative appendices (epidemiology and differential diagnosis) and two normative appendices (immune-detection and etiological examination) are attached. The Criteria provides the technical reference for diagnosis of kala-azar in medical institutions and disease control institutions. Combined with the current epidemic situation of kala-azar in China, this paper interprets the main contents of the Diagnostic Criteria for Kala-azar (WS 258-2006), so as to promote its learning and implementing.

Notch signaling modulates proliferative vitreoretinopathy via regulating retinal pigment epithelial-to-mesenchymal transition.

Elevated Notch signaling has been verified in a large range of fibrotic diseases developed in the kidney, liver, and lung, inducing the development of the epithelial-mesenchymal transition (EMT). The aim of this study was to observe the involvement of Notch signaling in the EMT of retinal pigment epithelial (RPE) cells and the pathogenesis of proliferative vitreoretinopathy (PVR). In vitro cultivated human RPE cells (ARPE-19) were treated with 10 ng/mL transforming growth factor (TGF)-ß1 for 24, 48, and 72 h. The expression levels of ZO-1, α-SMA, vimentin, Notch1 intracellular domain (NICD1), and Hes-1 were evaluated with quantitative real-time polymerase chain reaction (qRT-PCR), immunofluorescence staining or Western blot. TGF-ß1 induced EMT and the activation of Notch signaling in ARPE-19 cells. To examine the effect of Notch inhibition on TGF-ß1-induced EMT and PVR formation, ARPE-19 cells were preincubated with γ-secretase inhibitor LY411575 before TGF-ß1 treatment. Mouse PVR model was used for in vivo study. ARPE-19 cells were injected intravitreously with or without the LY411575 to examine the effect of Notch inhibition on PVR formation. LY411575 significantly attenuated EMT by inhibiting the Notch signaling activation in vitro. PVR was induced by intravitreal injections of ARPE-19 cells, while LY411575 inhibited mouse PVR formation in vivo. Notch signaling plays a critical role in TGF-ß1-induced EMT in vitro and mice PVR model, which provides a novel insight into the pathogenesis of PVR. The specific inhibition of Notch signaling by γ-secretase inhibitor may provide a new approach for the prevention of PVR.

Cloning and Expression Analysis of One Gamma-Glutamylcysteine Synthetase Gene (Hbγ-ECS1) in Latex Production in Hevea brasiliensis.

Rubber tree is a major commercial source of natural rubber. Latex coagulation is delayed by thiols, which belong to the important type of antioxidants in laticifer submembrane, and is composed of glutathione (GSH), cysteine, and methionine. The rate-limiting enzyme, γ-ECS, plays an important role in regulating the biosynthesis of glutathione under any environment conditions. To understand the relation between γ-ECS and thiols and to correlate latex flow with one-time tapping and continuous tapping, we cloned and derived the full length of one γ-ECS from rubber tree latex (Hbγ-ECS1). According to qPCR analysis, the expression levels of Hbγ-ECS1 were induced by tapping and Ethrel stimulation, and the expression was related to thiols content in the latex. Continuous tapping induced injury, and the expression of HbγECS1 increased with routine tapping and Ethrel-stimulation tapping (more intensive tapping). According to expression in long-term flowing latex, the gene was related to the duration of latex flow. HbγECS1 was expressed in E. coli Rosetta using pET-sumo as an expression vector and the recombinant enzyme was purified; then we achieved 0.827 U/mg specific activity and about 66 kDa molecular weight. The present study can help us understand the complex role of Hbγ-ECS in thiols biosynthesis, which is influenced by tapping.

[Cloning, expression and serological evaluation of H3 protein from Echinococcus granulosus].

OBJECTIVE: To clone and express the basement membrane specific heparan sulfate proteoglycan core protein (H3), and to evaluate its effect in detection of human cystic echinococcosis (CE). METHODS: The H3 gene immunoscreened from the cDNA library was cloned into pGEX-4T vector. The recombinant plasmid pGEX-3X-AgB8/3 was transformed into Escherichia coli BL21 strains and induced by isopropyl-ß-D-thiogalactopyranoside (IPTG). Then the expressed recombinant protein was purified by affinity chromatography and its effect in the detection of CE was evaluated by ELISA. Meanwhile, the effects of H3 and two antigens that the research group prepared before (purified HCF and rAgB8/2) in CE detection were compared. RESULTS: The plasmid pGEX-4T-H3 was successfully constructed and H3 was successfully expressed in prokaryotic cells. The sensitivities of the recombinant H3, purified HCF and rAgB8/2 in CE detection were 84.0% (68/81), 90.1% (73/81) and 77.8% (63/81) respectively, and there was no statistical difference among them (χ2 = 4.58, P > 0.05). The cross reactions of recombinant H3 with the sera of the patients with CE, cysticercosis and schistosomiasis were 63.3% (19/30), 16.7% (5/30) and 5.0% (1/20) respectively, and the cross reaction was 0 with the sera of healthy people. The specificities of recombinant H3, purified HCF, and rAgB8/2 were 80.8% (105/130), 71.5% (93/130) and 82.3% (107/130) respectively, and there were no statistical difference among them (χ2 = 5.71, P > 0.05). CONCLUSIONS: The recombinant H3 is a potential diagnostic antigen for CE detecting.

Protein phosphatase 2A is involved in the tyrosine hydroxylase phosphorylation regulated by α-synuclein.

α-Synuclein (α-Syn) plays a crucial role in the pathophysiology of Parkinson's disease (PD), the degeneration of dopaminergic neurons. Previous studies have shown that α-Syn regulates dopamine synthesis by binding to and inhibiting tyrosine hydroxylase (TH). In neurons, protein phosphatases (PPs) play a prominent role in directing signaling toward survival or degeneration. This study was to re-evaluate whether α-Syn could regulate the tyrosine hydroxylase phosphorylation by protein phosphatase-2A (PP2A) in dopaminergic MN9D cells and cortex neurons. Our data demonstrated for the first time that α-Syn stimulates PP2A activity and reduces phosphorylation of TH through regulating the methylation of PP2A in dopaminergic MN9D cells and primary cortex neurons. Increased PP2A activity and reduced phosphorylation of PP2A at Y307 (inactive form of PP2A) were observed in α-Syn overexpression dopaminergic cells (Syn) and primary cortex neurons, and the TH phosphorylation relieved by enhancing PP2A methylation in Syn group could be abated by using PP inhibitors, okadaic acid (OKA). OKA could reduce the cell damage and cell apoptosis induced by α-Syn. Thus our findings may provide an insight into the complicated pathogenesis of PD as well as some clues to the development of novel therapeutic strategies targeting at PP2A.

Risk factors for pancreatic infection in patients with severe acute pancreatitis: an analysis of 163 cases.

OBJECTIVE: We aimed to identify the risk factors for predicting pancreatic infection in patients with severe acute pancreatitis (SAP). METHODS: In all, 163 patients with SAP were included and divided into two groups based on the presence or absence of pancreatic infection. Their demographic and clinical characteristics, laboratory examination results, complications and treatment modalities were collected from their medical records. Variables were initially screened by univariate analysis and those with statistical significance were then filtered by multivariate analysis to determine the independent risk factors for pancreatic infection in SAP. RESULTS: Patients having SAP with pancreatic infection had a lower partial pressure of arterial carbon dioxide (PaCO2 ), peripheral white blood cell count and alkaline phosphatase levels, together with a higher computed tomography severity index (CTSI) than those without pancreatic infection, while their lactate dehydrogenase (LDH) levels and blood urea nitrogen were much higher. Pancreatic infection was also more common in patients receiving late fluid resuscitation than in those receiving early fluid resuscitation. Multivariate analyses revealed that increased LDH level, high CTSI, delayed fluid resuscitation and hypoxemia were independent risk factors for pancreatic infection in SAP. The sensitivity, specificity, positive and negative predictive values for a model combining the parameters in predicting pancreatic infection were 84%, 97%, 88% and 96%, respectively, with a cut-off value of 0.393, and the area under the receiver operating characteristic curve was 0.923. CONCLUSION: Increased LDH, high CTSI, delayed fluid resuscitation and hypoxemia are independent risk factors for predicting pancreatic infection in patients with SAP.

Evaluation of visualized area percentage assessment of cleansing score and computed assessment of cleansing score for capsule endoscopy.

BACKGROUND/AIM: The purpose of this study was to evaluate the clinical significance of visualized area percentage assessment of cleansing score (AAC) and computed assessment of cleansing score (CAC) of these two small bowel cleanliness scores systems for capsule endoscopy (CE). MATERIALS AND METHODS: The reliability and consistency of the AAC and CAC scores were evaluated by comparing the scores by two examiners (one expert, one without any training in CE). Reliability was determined using the intraclass correlation coefficient (ICC) and consistency was determined using the kappa statistic. RESULTS: The inter-observer agreement was excellent for both the AAC and CAC scores. For AAC, the ICC was 0.791 (95% confidence interval: 0.677-0.868), and for CAC the ICC was 1.000. Using 1.5 as the cutoff, there was no significant difference between AAC and CAC results by the expert examiner (kappa = 0.756, P = 0.000) or the non-expert examiner (kappa = 0.831, P = 0.000). Evaluation of small bowel cleanliness using AAC took 15-30 min, and evaluation using CAC took about 2-3 min. The overall adequacy assessment (OAA) using the AAC was not significantly different between the two examiners (χ2 = 0.586, P = 0.444). There were also no significant differences between the OAA using the AAC and the OAA using the CAC by the expert examiner (χ2 = 1.730, P = 0.188) or the non-expert examiner (χ2 = 1.124, P = 0.289). CONCLUSION: Both of these scores for assessment of small bowel cleanliness can be useful in clinical practice, but the CAC is simpler to use.