Whole transcriptome sequencing for revealing the pathogenesis of sporotrichosis caused by Sporothrix globosa.

This study aimed to investigate the molecular mechanism of sporotrichosis and identify possible novel therapeutic targets. Total RNA was extracted from skin lesion samples from sporotrichosis patients and used to construct a long-chain RNA transcriptome library and miRNA transcriptome library for whole transcriptome sequencing. The differentially expressed genes (DEGs) between the groups were identified, and then Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and Gene Set Enrichment Analysis enrichment analyses were performed based on the DEGs. An lncRNA-miRNA-mRNA ceRNA network was constructed. The expressions of JAK/STAT pathway-related proteins were detected in the patient and control tissues using RT-qPCR and Western blot analysis. Enrichment analysis showed that the DEGs were mainly enriched in various infectious diseases and immune response-related signaling pathways. Competing endogenous RNA network analysis was performed and identified the hub lncRNAs, miRNAs, and mRNAs. Compared with the control group, the mRNA expressions of SOCS3, IL-6, and JAK3 were significantly upregulated, while the expression of STAT3 did not change significantly. Also, the protein expressions of SOCS3, IL-6, JAK3, and STAT3, as well as phosphorylated JAK3 and STAT3, were significantly upregulated. We identified 671 lncRNA DEGs, 3281 mRNA DEGs, and 214 miRNA DEGs to be involved in Sporothrix globosa infection. The study findings suggest that the JAK/STAT pathway may be a therapeutic target for sporotrichosis.

Itraconazole therapy for infant hemangioma: Two case reports.

BACKGROUND: Infantile hemangiomas (IHs) are the most common childhood benign tumors, showing distinctive progression characteristics and outcomes. Due to the high demand for aesthetics among parents of IH babies, early intervention is critical in some cases. ß-Adrenergic blockers and corticosteroids are first-line medications for IHs, while itraconazole, an antifungal medicine, has shown positive results in recent years. CASE SUMMARY: In the present study, itraconazole was applied to treat two IH cases. The therapeutic course lasted 80-90 d, during which the visible lesion faded by more than 90%. Moreover, no obvious side effects were reported, and the compliance of the baby and parents was desirable. CONCLUSION: Although these outcomes further support itraconazole as an effective therapeutic choice for IHs, large-scale clinical and basic studies are still warranted to improve further treatment.

Comparison of leaf transcriptome in response to Rhizoctonia solani infection between resistant and susceptible rice cultivars.

BACKGROUND: Sheath blight (SB), caused by Rhizoctonia solani, is a common rice disease worldwide. Currently, rice cultivars with robust resistance to R. solani are still lacking. To provide theoretic basis for molecular breeding of R. solani-resistant rice cultivars, the changes of transcriptome profiles in response to R. solani infection were compared between a moderate resistant cultivar (Yanhui-888, YH) and a susceptible cultivar (Jingang-30, JG). RESULTS: In the present study, 3085 differentially express genes (DEGs) were detected between the infected leaves and the control in JG, with 2853 DEGs in YH. A total of 4091 unigenes were significantly upregulated in YH than in JG before infection, while 3192 were significantly upregulated after infection. Further analysis revealed that YH and JG showed similar molecular responses to R. solani infection, but the responses were earlier in JG than in YH. Expression levels of trans-cinnamate 4-monooxygenase (C4H), ethylene-insensitive protein 2 (EIN2), transcriptome factor WRKY33 and the KEGG pathway plant-pathogen interaction were significantly affected by R. solani infection. More importantly, these components were all over-represented in YH cultivar than in JG cultivar before and/or after infection. CONCLUSIONS: These genes possibly contribute to the higher resistance of YH to R. solani than JG and were potential target genes to molecularly breed R. solani-resistant rice cultivar.

Manipulation of the rice L-galactose pathway: evaluation of the effects of transgene overexpression on ascorbate accumulation and abiotic stress tolerance.

Ascorbic acid (AsA) is the most abundant water-soluble antioxidant in plants, and it plays a crucial role in plant growth, development and abiotic stress tolerance. In the present study, six key Arabidopsis or rapeseed genes involved in AsA biosynthesis were constitutively overexpressed in an elite Japonica rice cultivar. These genes encoded the GDP-mannose pyrophosphorylase (GMP), GDP-mannose-3',5'-epimerase (GME), GDP-L-galactose phosphorylase (GGP), L-galactose-1-phosphate phosphatase (GPP), L-galactose dehydrogenase (GDH), and L-galactono-1,4-lactone dehydrogenase (GalLDH). The effects of transgene expression on rice leaf AsA accumulation were carefully evaluated. In homozygous transgenic seedlings, AtGGP transgenic lines had the highest AsA contents (2.55-fold greater than the empty vector transgenic control), followed by the AtGME and AtGDH transgenic lines. Moreover, with the exception of the AtGPP lines, the increased AsA content also provoked an increase in the redox state (AsA/DHA ratio). To evaluate salt tolerance, AtGGP and AtGME transgenic seedlings were exposed to salt stress for one week. The relative plant height, root length and fresh weight growth rates were significantly higher for the transgenic lines compared with the control plants. Altogether, our results suggest that GGP may be a key rate-limiting step in rice AsA biosynthesis, and the plants with elevated AsA contents demonstrated enhanced tolerance for salt stress.

[Molecular investigation of a possible case of HIV transmission after a blood transfusion].

OBJECTIVE: A molecular technique based on quasispecies analysis for tracing postexposure HIV transmission was applied in an investigation of a possible case of HIV transmission after blood transfusion. METHODS: Sixteen plasma specimens were collected from 3 HIV infections (T1-T3) involved in a possible HIV transmission chain and 13 HIV/AIDS (C1-C13) controls. The RNAs were extracted and then amplified by RT-PCR, the PCR products were cloned and sequenced.BioEdit 6.0.7 and MEGA 4.0 software were used to analyze gene sequences, calculate gene dispersion ratio and construct phylogenetic tree. RESULTS: The sequences of 13 specimens were successfully obtained.The HIV strains from T1, T2 and T3 were CRF07_BC recombinants, those from 5 out of the 6 controls lived in the same city with T2 and T3 were CRF07_BC recombinants as well, while those from 4 controls living in the same city with T1 were CRF01_AE recombinants. Compared with the clone sequences from T1, the mean gene dispersion ratio of T2 was the least (2.0%), followed by C12 (2.8%) , T3 (2.9%) and others. The phylogenetic tree showed that all clones from T1, T2, T3 and C12 might cluster together,and implied that the direction of HIV transmission was from T3 to T2, and then to T1. CONCLUSION: The results support the possible epidemiological clue that HIV was transmitted from T3 to T2, and then to T1, indicating that molecular epidemiological investigation could provide more direct evidence for tracing postexposure HIV transmission.

Effects of N-stearoyl-L-tyrosine on the hippocampal ubiquitin-proteasome system in rats with chronic cerebral hypoperfusion.

OBJECTIVES: Chronic cerebral hypoperfusion (CCH) leads to neurodegeneration and cognitive impairment. Ubiquitinated protein aggregates are commonly present in neurodegenerative disorders and are believed to cause neuronal degeneration. Here, we investigated the effects of N-stearoyl-L-tyrosine (NSTyr) on the hippocampal ubiquitin-proteasome system (UPS) in rats with CCH. METHODS: After induction of CCH, NSTyr was intraperitoneally administered daily for 3 months. Protein aggregation was analyzed by ethanolic phosphotungstic acid (EPTA) electron microscopy (EM), immunogold EM, laser-scanning confocal microscopy, and Western blot. Proteasome peptidase activity was measured by peptidase activity assays. RESULTS: By using EPTA EM, immunogold EM and high-resolution laser-scanning confocal microscopy, we found that CCH resulted in the accumulation of ubiquitinated protein aggregates in rat hippocampal CA1 neurons. Western blot revealed that the levels of free ubiquitin were significantly reduced and that the levels of ubiquitinated proteins were markedly increased in the hippocampus of CCH rats. Direct activity measurements demonstrated that proteasome peptidase activity in the hippocampal region of rats was decreased after CCH induction. In the hippocampal tissue of CCH rats treated with NSTyr, however, ubiquitinated protein aggregates decreased and proteasome peptidase activity increased. DISCUSSION: These data indicate that NSTyr may exert protective effects on rat hippocampal UPS function via endogenous regulation.

Increased α-tocotrienol content in seeds of transgenic rice overexpressing Arabidopsis γ-tocopherol methyltransferase.

Vitamin E comprises a group of eight lipid soluble antioxidant compounds that are an essential part of the human diet. The α-isomers of both tocopherol and tocotrienol are generally considered to have the highest antioxidant activities. γ-tocopherol methyltransferase (γ-TMT) catalyzes the final step in vitamin E biosynthesis, the methylation of γ- and δ-isomers to α- and ß-isomers. In present study, the Arabidopsis γ-TMT (AtTMT) cDNA was overexpressed constitutively or in the endosperm of the elite japonica rice cultivar Wuyujing 3 (WY3) by Agrobacterium-mediated transformation. HPLC analysis showed that, in brown rice of the wild type or transgenic controls with empty vector, the α-/γ-tocotrienol ratio was only 0.7, much lower than that for tocopherol (~19.0). In transgenic rice overexpressing AtTMT driven by the constitutive Ubi promoter, most of the γ-isomers were converted to α-isomers, especially the γ- and δ-tocotrienol levels were dramatically decreased. As a result, the α-tocotrienol content was greatly increased in the transgenic seeds. Similarly, over-expression of AtTMT in the endosperm also resulted in an increase in the α-tocotrienol content. The results showed that the α-/γ-tocopherol ratio also increased in the transgenic seeds, but there was no significant effect on α-tocopherol level, which may reflect the fact that γ-tocopherol is present in very small amounts in wild type rice seeds. AtTMT overexpression had no effect on the absolute total content of either tocopherols or tocotrienols. Taken together, these results are the first demonstration that the overexpression of a foreign γ-TMT significantly shift the tocotrienol synthesis in rice, which is one of the world's most important food crops.

A dural arteriovenous fistula in cavernous sinus developed from viral meningitis.

Although hormonal influences, inflammation, trauma, sinus thrombosis, venous hypertension, and congenital origin have been proposed as sources of dural arteriovenous fistulas (DAVFs) in cavernous and sigmoid sinuses, the etiology of these lesions remains controversial. We present a case with a cavernous sinus DAVF developed from viral meningitis which has not been previously described. A 24-year-old male was admitted to our institute because of periorbital pain, decreased vision, pulsatile tinnitus, chemosis, and exophthalmos on the right side after he had suffered viral meningitis four months before. Cerebral angiography demonstrated a cavernous sinus DAVF, which was successfully obliterated with several platinum coils using a transvenous approach. The viral meningitis most likely caused the inflammation, that may be responsible for the occurrence of the cavernous sinus DAVF. Prompt treatment for inflammation may help to prevent the development of DAVFs.

[Comparison of three HIV antibody confirmatory assay kits in confirming early HIV infection].

OBJECTIVE: This study was to compare the performance of three HIV antibody confirmatory assay kits in confirming early HIV infection. METHODS: Five HIV antibody-positive plasma specimens were ten-fold serially diluted and then detected by ELISA. The above diluted specimens were detected with the following three HIV antibody confirmatory assay kits to analyze their sensitivity, including Wantai-RIBA (Recombinant immunoblot assay, Beijing Wantai Biological Pharmacy, China), MP-WB (HIV Blot 2.2 WB, MP Biomedicals Asia Pacific Pte. Ltd., Singapore) and INNO-LIA (INNO-LIA(TM) HIV I/II Score, Innogenetics N.V., Belgium), respectively. These kits were further used to detect 48 ELISA-reactive specimens from 11 sets of HIV seroconversion specimens (a total of 48 samples) which were previously detected as HIV antibody-positive by ELISA. RESULTS: When 5 samples were diluted to 100 fold, Wantai-RIBA still can detect them positive. Among the 48 HIV antibody-positive specimens detected with ELISA, the confirmation positive rate for Wantai-RIBA, MP-WB and INNO-LIA were 97.92% (47/48), 81.25% (39/48) and 91.67% (44/48), respectively. There was statistically significant difference between the confirmatory results of Wantai-RIBA and MP-WB (χ(2) = 6.13, P < 0.05), as well as between those of INNO-LIA and MP-WB (χ(2) = 5.48, P < 0.05); however, there was no statistically significant difference between those of Wantai-RIBA and INNO-LIA (χ(2) = 1.33, P > 0.05). For other six HIV seroconversion panels containing indeterminate specimens, the average seroconversion period of time for Wantai-RIBA, MP-WB and INNO-LIA were 0.7, 13.3 and 3.7 days, respectively. CONCLUSION: Compared with MP-WB, Wantai-RIBA and INNO-LIA could reduce the window period to confirm early HIV infection.

Establishment of a novel hemodynamic cerebral ischemia model of atherosclerotic rabbit.

BACKGROUND: Atherosclerosis is the most common cause of ischemic stroke. Until now, there has been no ideal animal model for studying the hemodynamic ischemia caused by atherosclerosis in posterior circulation. OBJECTIVE: This study was conducted to establish a cerebral ischemic model in an atherosclerotic rabbit by subclavian artery (SA) ligation or embolization, emphasizing the role of hemodynamics. MATERIALS AND METHODS: Thirty-two male New Zealand rabbits (weight, 3.5 to 4.0 kg; provided by Animal Laboratory of Tongji Hospital of Tongji University) were randomly divided into four groups. All the groups, except the controlled group, were fed with high lipid diet for three months and two of the groups were submitted for right SA ligation or left SA embolization and the remaining one group to non-operated group. RESULTS: A SA steal syndrome was established successfully with delayed visualization and prolonged circulation time in the posterior circulation. The circulation time was different when SA was ligated or embolized (P < 0.001), and also there was statistically significant difference between controlled and non-operated groups when angiography of either, left and right, VA was performed (P = 0.009, P = 0.003, respectively). However, there was no statistical significant difference between the ligation and embolization group either before or after the procedure (ligation or embolization) (P = 0.402, P = 0.068, respectively). In the control and non-operated group, angiography of either left or right VA showed no statistically significant difference (P = 0.285, P = 0.197, respectively). In the groups fed with high lipid diet for three months, atherosclerotic plaques were found in the common carotid arteries (CCAs), aortic arch (AAs) and variably in the internal carotid arteries (ICAs). CONCLUSIONS: This model could, to some extent, simulate the clinical hemodynamic state of atherosclerotic stroke. and it is an ideal model for studying the hemodynamic cerebral ischemia in posterior circulation.

Characterization of expression of the OsPUL gene encoding a pullulanase-type debranching enzyme during seed development and germination in rice.

Starch-debranching enzymes (DBEs) are key enzymes involved in starch metabolism in cereals, having a dual function, in both starch synthesis and degradation. However, their precise roles in this pathway, particularly their expression profiles, remain unclear. In the present study, we performed a quantitative real-time PCR (Q-PCR) analysis of the expression pattern of the OsPUL gene encoding a pullulanase-type DBE in different tissues as well as in seeds at different developmental stages. The results showed that this gene was expressed only in seeds. In addition, the 1177-bp OsPUL promoter sequence was cloned, and some endosperm-specific motifs such as the GCN4 and AACA motifs were observed to exist in this region. The promoter was then fused with the GUS reporter gene and its expression was carefully investigated in transgenic rice. The data from both histochemical and fluorometric analyses showed that the OsPUL promoter was capable of driving the target gene to have a high level of endosperm-specific expression. The OsPUL gene maintained a relatively high expression level during the entire period of seed development, and peaked in the middle and late stages. This observation was very consistent with that of the endogenous transcription analysis by Q-PCR. Furthermore, the seed germination experiment showed that the OsPUL promoter actively functions in the late stage of seed germination. The expression of the OsPUL gene was maintained at a significant level during the entire grain filling period and in the late stage of seed germination, which coincided with its involvement in starch anabolism and catabolism.

Proteomic analysis of 'hybrid necrosis' in wheat (Triticum aestivum) leaves.

Wheat hybrid necrosis has been genetically characterised for many years, but the specific gene(s) and the protein products involved in the processes remains unknown. In this study, protein expression in the base (B), mid (M) and tip (T) segments of the FL-2 leaves of a necrotic hybrid, PZF1 and its parents, Pan555 and Zheng891, was analysed and compared using a high throughput proteomic approach. Twenty-three protein spots, with significant variations in intensity across the necrotic leaf segments, were analysed by MALDI-TOF-MS, of which, 18 were matched to protein accessions in the NCBI database. Several of these proteins are enzymes involved in the methylation cycle, including AdoHcy hydrolase, AdoMet synthase 3 and methionine synthase 1; AdoHcy hydrolase was downregulated sharply in M and T, and AdoMet synthase 3 and methionine synthase 1 were upregulated gradually from M to T. This result suggests that methylation-associated processes, including epigenetic mechanisms, may play a role in the initiation and development of hybrid necrosis. Several energy cycle-associated proteins and cytoprotective proteins were also differentially expressed across the leaf segments, suggesting their direct association with or possible involvement in the necrotic processes. The significant imbalance of a heat-shock protein, a transposon protein and a RNA- and ssDNA-binding protein also makes these proteins potential molecular components in the necrotic processes.

Epidemiology, clinical and laboratory characteristics of currently alive HIV-1 infected former blood donors naive to antiretroviral therapy in Anhui Province, China.

BACKGROUND: Unregulated commercial blood/plasma collection among farmers occurred between 1992 and 1995 in central China and caused the second major epidemic of human immunodeficiency virus type 1 (HIV-1) infection in China. It is important to characterize HIV-1-infected former blood donors and to study characteristics associated with disease progression for future clinical intervention and vaccine development. METHODS: A cross-sectional study was performed on HIV-1-infected former blood donors (FBDs) and age-matched HIV-seronegative local residents. Demographic, epidemiologic, clinical and key laboratory data were collected from all study participants. Both unadjusted and adjusted multivariate linear regressions were employed to analyze the association of the decrease of CD4(+) T-cell counts with other characteristics. RESULTS: Two hundred and ninety-four HIV-1-infected FBDs and 59 age-matched HIV-seronegative local residents were enrolled in this study. The unregulated blood/plasma collection occurred more than a decade (10.8 - 12.8 years) ago, which caused the rapid spread of HIV-1 infection and the high prevalence of co-infection with hepatitis C virus (HCV, 89.5%); hepatitis B virus (HBV) co-infection was observed in only 11 HIV(+)participants (3.7%). Deterioration in both clinical manifestation and laboratory parameters and increase of viral loads were observed in parallel with the decrease of CD4(+) T-cell counts. The decrease of total lymphocyte counts (P < 0.001) and hemoglobin levels (P < 0.001) and the appearance of dermatosis (P = 0.03) were observed in parallel with the decrease of CD4(+) T-cell counts whereas viral loads (P < 0.001) and CD8(+) T-cell counts (P = 0.01) were inversely associated with CD4(+) T-cell counts. CONCLUSIONS: Co-infection with HCV but not HBV is highly prevalent among HIV-1-infected FBDs. CD4(+) T-cell counts is a reliable indicator for disease progression among FBDs. Total lymphocyte counts, hemoglobin level and appearance of dermatosis were positively associated with CD8(+) T-cell counts and viral loads were inversely associated with the decreased CD4(+) T-cell counts.

[Effect of duration and temperature of sample preservation on the result of peripheral blood CD4+ and CD8+ T lymphocyte count in HIV/AIDS patients].

BACKGROUND: By analyzing the CD4+ and CD8+ T lymphocyte count of whole blood from HIV/AIDS patients, which were stored at different temperatures for various durations, the authors studied the ideal preserving condition for whole blood and processed, in a purpose of guaranteeing the accuracy of clinical testing of CD4+ and CD8+ T lymphocyte count. METHODS: Blood from 34 HIV carriers/AIDS patients, were kept at 4 degrees C for 2, 24, 48, or 72 h, and tested for CD4+ and CD8+ T lymphocyte count using cytometric analysis. Part of the blood was processed, and kept at degrees C or room temperature for 2, 24, 48, or 72 h, then tested for CD4+ and CD8+ T lymphocyte count. The results were compared statistically in parallel. RESULTS: Whole blood and processed samples preserved at degrees C showed no statistical difference in CD4+ T lymphocyte count among different preserving durations (P greater than 0.05), but CD8+ T lymphocyte counts were significantly different at 72 h (P less than 0.05). Processed samples at 72 h were significantly different in CD4+ T lymphocyte count(P less than 0.05), and significantly different in CD8+ T lymphocyte count at 24 h (P less than 0.05). At room temperature, samples at different duration were not significantly different in CD4+ T lymphocyte count, but significantly different in CD8+ T lymphocyte count at 48 and 72 h (P less than 0.05). CONCLUSION: There were stable results for performing analysis of the CD4+ and CD8+ T lymphocyte count of the anticoagulated blood within 48 h. At room temperature, there were stable results for performing the analysis of CD4+ and CD8+ T lymphocyte count of processed samples within 24 h. Between 24 h and 48 h, although CD4+ count was stable, CD8+ count showed significant changes, so the ratio of CD4 to CD8 changed accordingly.

[An epidemiological study on sexual transmission of human immunodeficiency virus among pre-marital group in Yining city, Xinjiang].

OBJECTIVE: To study the human immunodeficiency virus (HIV) status through heterosexual transmission in Yining city and to provide information on effective intervention measures. METHODS: Cohort of HIV sero-discordant couples identified from 1997 to 2000 was formed. Proportional risk model was used to analyze the time of HIV sero-conversion and the related factors. All the recruiters were under informed consent. RESULTS: Through following on 22 sero-discordant couples, we found that the incidence density (ID) of HIV sero-conversion was 32.49/100 person-year (PY) with 33.74/100 PY for women. In the proportional hazard model, the course of sero-conversion was only 2.43 years and the frequency of sexual contact was statistically significant (>or= 3 times/week vs. < 3 time/week: RR = 1.984, 95% CI: 1.045 - 3.767), indicating this factor was related to the hazard of HIV sero-conversion. However, the viral load of HIV infections has no such effect on HIV sero-conversion of their spouses. In addition, the ratio of CD4(+)/CD8(+) was lower in spouses of HIV sero-conversion than that in spouses of HIV non-sero conversion (t test: t = 4.77, P < 0.01). CONCLUSION: In order to control HIV transmission among general population, we suggested that HIV/AIDS counseling and testing be developed for pre-marital people in the region with high HIV prevalence.