[Diagnostic value of heparin-binding protein in patients with silicosis complicated by severe infection].

Objective: To explore the application value of heparin binding protein (HBP) in the diagnosis of severe infection in patients with silicosis. Methods: A prospective study was conducted on 150 patients with silicosis in the pneumoconiosis department of the General Hospital of Xuzhou Mining Group from January 2017 to March 2018. Among them, 100 were severely infected with silicosis and 50 were non-infected with silicosis. 30 patients were selected in the same period of physical examination as the control group. HBP, C-reactive protein (CRP) , procalcitonin(PCT) , white blood cell count (WBC) , neutrophil percentage, and absolute neutrophil count(ANC) were detected in all participants. Using the receiver operating characteristic curve(ROC) to analyze the diagnostic value of indicator above in patients with different stages of severe silicosis infection. Results: Plasma HBP levels in patients with severely infected silicosis group[(50.39±35.64) ng/ml] were higher than those in the non-infected group[(10.71±1.47) ng/ml] and the control group[(9.24±1.83) ng/ml] (P<0.05) , and with the increase of silicosis stages, there is an increasing trend (P<0.05). The ROC curve showed that the AUC of HBP in the patients with severe silicosis in the first, second, and third stages were 0.932, 0.977, and 0.964, which were higher than those of WBC, CRP, and PCT. Correlation analysis showed that HBP was positively correlated with WBC, CRP and PCT (r=0.711, 0.359, 0.729, P<0.01). Conclusion: HBP has high diagnostic efficacy in the diagnosis of severe infections in patients with silicosis, which may become a clinical screening indicator for severe infections in patients with silicosis and an auxiliary examination indicator for the stage of silicosis patients.

[Expression of mRNA and protein of Klotho gene in placental tissue of macrosomia and its relationship with birth weight of neonates].

OBJECTIVE: To explore the the expression of Klotho mRNA and protein in placenta of macrosomia and its relationship with the birth weight of neonates. METHODS: The cases were from November 2014 to March 2015 in Shengjing Hospital of China Medical University, divided into 4 groups: the gestational diabetes with macrosomia group (GM), the gestational diabetes with normal birth weight group (GN), the normal pregnancy with macrosomia group (NM) and the normal pregnancy with normal birth weight group (NN). Klotho mRNA and protein expression in the placenta were detected by immunohistochemistry SP method, real-time fluorescent quantitative PCR and western blot, respectively, and were compared among the 4 groups. RESULTS: (1) Immunohistochemical detection showed the positive rate of Klotho protein was significantly higher in the placenta of GM (93%,28/30) than in the GN (73%,22/30; P<0.05). The positive rate was significantly higher in the placenta of NM (97%,29/30) than in the NN (80%,24/30; P<0.05). (2) Real-time fluorescent quantitative PCR showed the Klotho mRNA expression was significantly higher in the placenta of GM (4.3 ± 3.1) than in the GN (2.1 ± 2.4; P<0.05). The Klotho mRNA expression was also significantly higher in the placenta of NM (4.8± 3.4) than in the NN (2.6± 3.3; P<0.05). (3) Western blot showed the Klotho protein expression was significantly higher in the placenta of GM (1.27±0.90) than in the GN (0.64±0.24; P<0.05). It was also significantly higher in the placenta of NM (2.51±3.52) than in the NN (0.77±0.37; P<0.05). (4) There were no significant differences in the expression of Klotho mRNA and protein between GM and NM, GN and NN (P>0.05). CONCLUSIONS: The up-regulation of Klotho gene may be associated with macrosomia. The relationship is not affected by the complication of gestational diabetes.

Screening of genes related to ovarian development in the swimming crab, Portunus trituberculatus, by suppression subtractive hybridization.

The swimming crab, Portunus trituberculatus, is an important marine animal and is widely cultured in China. In the present study, suppression subtractive hybridization was applied to identify the differentially expressed genes in the ovaries of mature and immature P. trituberculatus. One hundred and seventy six expressed sequence tag (ESTs) were identified, of which 100 were down-regulated, and 76 up-regulated. BLAST analysis identified 51 unigenes, of which 27 were down-regulated, and 24 up-regulated. Quantitative real-time reverse transcriptase polymerase chain reaction results indicated that the SSH technique is valuable in screening genes related to ovarian development. Genes identified in this study encoded proteins corresponding to a wide range of functions and included immune response protein, transcription initiation factor, metabolic proteins, chromosome, histone h3, ovarian development-related protein, and vitellogenin. In addition, 64 metabolic pathways were annotated in differentially expressed ESTs by using the Kyoto Encyclopedia of Genes and Genomes pathway. Four annotated pathways (oxidative phosphorylation, carbon metabolism, fatty acid degradation, and protein digestion and absorption) appeared to be involved in ovarian development. In ontology analysis, 5.83% of the cellular process genes in reverse subtraction cDNA library are involved in reproduction, and 5.88% involved in developmental process. In up-regulated genes, myosin II-expressed polehole-like protein; histone h3; ovigerous-hair stripping substance; peritrophin 48; and ovarian development-related protein appeared to be involved in ovarian development. Identification of differentially expressed genes in the mature and immature ovary of the swimming crab provides new insights for further studies on the mechanism underlying ovarian development in this species.

Molecular cloning and characterization of a novel Y-box gene from Sepiella maindroni.

Y-box proteins are a family of highly conserved nucleic acid binding proteins that interact with genome and transcription product to modulate the transcriptional and translational processes. In the present study, a complete mRNA of Y-box binding protein (designated SmYB) was obtained from Sepiella maindroni by amplification of flanking sequences. The full size of SmYB cDNA was 1502 bp, including 99 bp at the 5ꞌ untranslated region (UTR), a 3ꞌ UTR of 821 bp with a poly (A) tail, and an open reading frame of 582 bp, encoding a polypeptide of 193 amino acids with the predicted molecular weight of 16.48 kDa. The conserved cold-shock domain and two known RNA binding motifs identified in SmYB strongly suggested that SmYB was a new member of Y-box proteins. Quantitative real-time PCR was performed to examine the expression of SmYB mRNA in various tissues, embryos, and its temporal expression in liver after cold shock. The mRNA transcript of SmYB was detected in all examined tissues, with the highest expression level in testis and ovary. SmYB was abundant in early developmental stages of S. maindroni embryos but diminished in the late post-embryonic development. In addition, cold-shock treatment upregulated the transcription of SmYB mRNA in liver. These results demonstrated that SmYB is involved in embryonic development of S. maindroni and its tolerance to acute low temperatures.

mRNA expression profiles of heat shock proteins of wild and salinity-tolerant swimming crabs, Portunus trituberculatus, subjected to low salinity stress.

Challenged by the low salinity, 4 parts per thousand (4 ppt), for 72h, the survivals of swimming crabs (Portunus trituberculatus) were collected as the screened group (SG, tolerant to low salinity). Aiming at identifying the mechanism of low salinity tolerance, quantitative real-time PCR was employed to investigate the expression profiles of 4 HSP genes (HSP60, HSP70, HSP90-1, HSP90-2) in the hepatopancreas of wild (WG) and screened (SG) groups of P. trituberculatus exposed to low salinity (4 ppt). The results showed that 3 of the candidate genes (HSP60, HSP70, HSP90-1) exhibited similarly downregulated expression profiles in the first 3 h (P < 0.05), which became upregulated from 3 h to 72 h after being subjected to low salinity conditions. In contrast, the expression profile of the HSP90-2 gene was upregulated during the first 6 h for the WG, and during the first 12 h for the SG, after which it became downregulated. HSP90-1 and HSP90-2 were highly expressed at 12 h after low salinity challenge in the SG, but not the WG. The response of these 2 genes to salinity stress indicates their suitability as biomarkers to differentiate SG from WG crabs. The results indicate that HSP genes are involved in the adaptation of crabs to low salinity exposure, and that different HSPs have diverse functions in response to low salinity stress in P. trituberculatus. In addition, HSP expression in SG indicates that this group is more tolerant to low salinity conditions compared to WG.

Development of polymorphic expressed sequence tag-single sequence repeat markers in the common Chinese cuttlefish, Sepiella maindroni.

The common Chinese cuttlefish (Sepiella maindroni) is one of the popular edible cephalopod consumed across Asia. To facilitate the population genetic investigation of this species, we developed fourteen polymorphic microsatellite makers from expressed sequence tags of S. maindroni. The number of alleles at each locus ranged from 6 to 10 with an average of 7.9 alleles per locus. The ranges of observed and expected heterozygosity were from 0.615 to 0.962 and 0.685 to 0.888, respectively. Four loci were found deviated significantly from Hardy-Weinberg equilibrium. The polymorphism information content ranged from 0.638 to 0.833. These polymorphic microsatellite loci will be helpful for the population genetic, genetic linkage map, and other genetic studies of S. maindroni.

First Report of Pindo Palm Heart Rot Caused by Ceratocystis paradoxa in China.

On January 12th, 2012, a novel disease with an incidence of 50% was discovered in Pindo palm Butia capitata (Mart.) Becc from the Coconut Grant View Garden (19°33.137' N, 110°47.482' E) located in Wenchang, Hainan Province. Diseased leaflets at the base of the rotted heart leaves had reddish brown lesions; when the infection progressed, the leaves turned yellow and became blighted from the inner to the outer part of the crown. Once the growing point was destroyed, the entire tree ultimately died. Tissues from the edges of lesions from diseased leaflet samples were placed onto potato dextrose agar (PDA) and incubated at 25°C for 3 days. The color of colonies of five isolates obtained turned from white to black in 48 h. The optimum temperature for mycelium growth was from 20 to 30°C, and no growth occurred at temperatures higher than 40°C or lower than 5°C (n = 5). The cylindrical colorless to pale brown conidia were 7.5 to 17.5 µm long × 5.0 to 7.5 µm wide (n = 100); oval black chlamydospores were 12.5 to 22.5 × 7.5 to 15.0 µm (n = 100). The sequence (497 bp) of the internal transcribed spacer (ITS) region of the representative isolate BX3 (China Center for Type Culture Collection No. CCTCC AF2014002) was amplified using primer pair ITS1/ITS4 (GenBank Accession No. KF939052) and shared 99% sequence identity with Ceratocystis paradoxa strain xie331-4 (JQ039332). Based upon these biological characteristics and ITS sequence, this pathogen was identified as C. paradoxa (Dade) C. Moreau (anamorph Thielaviopsis paradoxa (de Seynes) Höhn.) (3). Pathogenicity tests were conducted on 8-cm-long sections of young leaflets excised from a 12-year-old pindo palm tree. One side of the midrib of 10 sections was wounded with a sterilized scalpel at the center and the other side was non-wounded, then a PDA plug (4 to 6 × 4 to 6 mm) from the edge of an actively growing colony of BX3 incubated for 3 days were inoculated onto each wounded or non-wounded site. As controls, plain PDA plugs were placed on wounded and non-wounded spots of another 10 sections following the above procedure. Pathogenicity was tested twice. Each inoculated section was then put into a 9-cm petri dish in which two filter papers (Φ = 9 cm) were placed and 8 ml of sterile water were added to maintain high humidity, and then all dishes were placed in a dark incubator at 25°C. After 5 days, typical symptoms developed only on the wounded points inoculated with mycelium plugs. C. paradoxa was re-isolated from the margins of the expanding lesions. C. paradoxa causing fruit rot of B. capitata was reported in Uruguay (2), but to our knowledge, there are no previous reports of this species in China or infecting leaves of B. capitata worldwide (1). We report here a new Ceratocystis disease on B. capitata, and it was named as pindo palm heart rot based on its symptoms. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases, Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , Feb 21, 2014. (2) V. Gepp et al. New Dis. Rep. 27:12, 2013. (3) F. Y. Yu et al. Plant Dis. 96:290, 2012.

First Report of Leaf Spot of Tea Oil Camellia (Camellia oleifera) Caused by Lasiodiplodia theobromae in China.

Tea oil camellia (Camellia oleifera Abel.), one of the most famous woody oil plants, is distributed and cultivated widely in central and southern China for its strong adaptability. In September 2013, tea oil camellia plants with severe leaf spots were observed in commercial production fields located in Wenchang, Hainan Province. Spots were initially chlorotic, became necrotic and black with a chlorotic halo, developing to cover the entire width of the leaves, and leading to leaf death. Isolations were performed by excising pieces of symptomatic leaves from the lesion margin, surface sterilized with 90% ethanol and 0.6% sodium hypochlorite, and then placed them on potato dextrose agar (PDA). Plates were incubated in a sterile chamber at 26 ± 2°C for 2 days. A fungus was consistently isolated on PDA from all 23 diseased leaf samples. Pure cultures were obtained by monosporic culture technique. After 2 to 3 days of incubation at 26 ± 2°C with a 12-h photoperiod, the fungus initially produced white colonies with dense aerial mycelia, which later turned black (6 to 7 days). The mycelium was fast spreading, branched, and septate. Pycnidia were black, globose, ostiolate, and produced in stroma on the medium surface after 28 days at the same culture conditions as above. Conidia were initially unicellular, subovoid, hyaline, thick-walled with granular content, and 19.8 to 28.9 × 11.5 to 15.7 µm (avg. 25.1 × 13.5 µm). Mature conidia were one-septate and dark brown with longitudinal striations. These observed morphological features suggested that the fungus possessed the same characteristics as previously described for Lasiodiplodia theobromae (Pat.) Griffon & Maubl (syn = Botryodiplodia theobromae) (2). For molecular identification, the ITS1-5.8S-ITS2 region and fragments of the ß-tubulin and elongation factor 1-alpha (EF1-α) genes were sequenced and BLASTn searches done in GenBank. Accession numbers of gene sequences submitted to GenBank were KF811055 for ITS region; KJ639047 for ß-tubulin; and KJ639048 for EF1-α. For all genes used, sequences were 99 to 100% identical to reference isolate CBS164.96 of L. theobromae reported in GenBank (NR_111174, EU673110, and AY640258). Hence, both morphological and molecular characteristics confirmed the fungus as L. theobromae. To confirm fungal pathogenicity, ten 1-year-old healthy plants of C. oleifera were inoculated with the fungus. Mycelial plugs (5 mm) taken from a 7-day-old colony growing on PDA were deposited on wounds with a sterilized knife on leaves and covered with moist cotton. Ten additional control plants were treated similarly but with sterile PDA plugs. Plants were maintained in a moist chamber at 26 ± 2°C for 3 days and then in a greenhouse at 25°C and 40% relative humidity. All the inoculated plants produced typical leaf spot symptoms 3 weeks after inoculation. The fungus was consistently re-isolated from all inoculated plants. Control plants did not show any symptoms. L. theobromae has been reported to cause cankers and dieback in a wide range of hosts and is common in tropical and subtropical regions of the world (1,2), but not previously reported causing disease on C. oleifera. To our knowledge, this is the first report worldwide of leaf spot of C. oleifera caused by L. theobromae. References: (1) S. Mohali et al. For. Pathol. 35:385, 2005. (2) E. Punithalingam. Page 519 in: CMI Descriptions of Pathogenic Fungi and Bacteria. Commonwealth Mycological Institute, Kew, Surrey, UK, 1976.

First Report of Burkholderia andropogonis Causing Bacterial Leaf Spot of Betel Palm in Hainan Province, China.

In May 2009, a severe bacterial disease of arecanut (Areca catechu L.) with an incidence of 100% was observed in a plantation of about 8,400 plants in Wenchang City, Hainan Province, China (19°47.171' N, 110°54.335' E). Symptoms consisted of small circular to elongated brown lesions, ranging from 1 to 105 mm in length and 1 to 21 mm in width, surrounded by yellow halos. White colonies, without fluorescent or diffusible pigments, were consistently recovered on King's B Medium plates from lesions surface-sterilized in 70% ethyl alcohol for 1 min. All isolates were gram-negative and each had a single, polar, sheathed flagellum. Isolates were identified as a Burkholderia sp. based on physiological and biochemical tests: oxidase and catalase positive, negative for arginine dihydrolase, gelatin hydrolysis and starch hydrolysis, and negative for acid production from levan (1,3). Sequences (approx. 1,400 bp each) of the 16S rRNA gene amplified from four isolates using primer pair 27F/1492R (2) (GenBank Accession Nos. JX415481, JX415479, JX415482, and JX415483) shared 99% sequence identity with that of Burkholderia andropogonis strain 6369 (DQ786951). Representative isolates Y11 (China General Microbiological Culture Collection Center No. CGMCC 1.12337), Y30 (CGMCC 1.12338), W15, and W20 were compared with B. andropogonis strain NCPPB No. 1012 and all caused a hypersensitive reaction on leaves of Nicotiana benthamiana. Isolate pathogenicity was tested twice with a total of three replications per isolate. Two young leaves each of 2-year-old arecanut plants were infiltrated with a bacterial suspension of 108 CFU/ml, then covered individually with plastic bags for 48 h, and incubated at 100% relative humidity with 16 h of daylight at 25°C by day and 8 h of darkness at 20°C by night. After 7 days, small water-soaked spots with yellow halos were observed and 60 days after inoculation, lesions developed similar to those caused by B. andropogonis in the field. Koch's postulates were fulfilled by reisolating bacteria from typical lesions on inoculated plants. These bacteria were identical to inoculated strains in colony morphology and sequences of the 16S ribosomal RNA gene. To our knowledge, this is the first report of B. andropogonis infection on betel in Hainan Province, mainland China. This disease was first reported in Taiwan, a province of China. Conditions of high humidity and high temperature support disease outbreaks and infection can result in severe economic losses. In 2012, this disease also appeared on a number of plantations located in other counties. As betel is, economically, the second most important crop in Hainan Province, measures should be required to control this disease, especially in typhoon seasons. References: (1) S. H. Hseu et al. Plant Pathol. Bull. 16:131, 2007. (2) D. J. Lane. In: E. Stackebrandt, et al. Nucleic acid techniques in bacterial systematics. John Wiley & Sons, Chichester, United Kingdom, pp. 115-175, 1991. (3) X. Li and S. H. De Boer. Plant Dis. 89:1132. 2005.

First Report of Stem Bleeding in Coconut Caused by Ceratocystis paradoxa in Hainan, China.

Stem bleeding of coconut was discovered in 2009 in Hainan, China. Affected trunk areas exhibited dark discoloration and a reddish brown or rust-colored liquid bleeding from different points. Stem tissues under the lesions rotted and became brownish yellow to black. Affected plants died within 3 to 4 months after stem symptoms first appeared. Stem bleeding of coconut is known to occur in production areas worldwide. The disease was first reported in Sri Lanka (1), caused severe damage to PB-121 hybrids in Indonesia (2), and is now known to occur in many other coconut-producing countries. However, to our knowledge, this is the first report of the disease in China. A fungus was isolated from lesion margins of diseased coconut trees. Colonies on potato dextrose agar (PDA) were white, became black 1 to 2 days later, and emitted a strong, fruity aroma. The fungus produced conidia, which were cylindrical, colorless to pale brown, and 6.9 to 14.9 × 3.1 to 6.0 µm, and oval, black chlamydospores that were 7.9 to 19.4 × 4.6 to 11.0 µm. The optimum temperature for mycelial growth ranged from 25 to 35°C and it did not grow at temperatures lower than 5°C or higher than 40°C. On the basis of these characteristics, the fungus was identified as Ceratocystis paradoxa (Dade) C. Moreau (anamorph Thielaviopsis paradoxa (de Seynes) Höhn). The internal transcribed spacer (ITS) region was amplified from genomic DNA with primers ITS1 and ITS4 and the PCR products were sequenced (GenBank Accession No. JQ039332). BLAST analysis showed 99% sequence similarity with C. paradoxa (GenBank Accession No. HQ248205.1). Pathogenicity of the fungus was tested by inoculating 10, 3-year-old coconut trees of the cv. green tall at the 12-leaf stage in the field. Agar plugs (5 mm in diameter) from the periphery of 7-day-old C. paradoxa colonies grown on PDA were placed on healthy trunks, rachis, and leaves, which were either wounded or unwounded. Wounds were made with a sterilized cork borer. Sites of the inoculations were wrapped with plastic tape to prevent desiccation; the experiment was repeated three times. Controls received plain PDA discs. Two weeks after inoculation, characteristic rusty brown lesions appeared only on wounded plants that were inoculated with the fungus. A brownish liquid oozed from the points of inoculation. Controls did not show signs of disease development. C. paradoxa was reisolated from the diseased tissues. Infection occurred on wounded sites only, suggesting that wounds may be required for infection. To prevent stem bleeding of coconut trees by C. paradoxa, vigilant cultural practices must be maintained to avoid causing wounds on the trees. References: (1) S. A. Alfieri. Plant Pathol. Circular No. 53. Florida Department of Agriculture Division of Plant Industry, 1967. (2) D. R. N. Warwick and E. E. M. Passos. Trop. Plant Pathol. 34:175, 2009.

MiR-150 suppressed cell viability, invasion and EMT via HMGA2 in oral squamous cell carcinoma.

OBJECTIVE: Oral squamous cell carcinoma (OSCC) accounts for 90% of head and neck cancers, and its 5-year overall survival is very poor. MiR-150 is usually downregulated and acts as tumor suppressor in multiple cancers. The aim of our study is to explore the functions of miR-150 in OSCC. PATIENTS AND METHODS: Expressions of miR-150 and HMGA2 mRNA in OSCC tissues and cells were analyzed by qRT-PCR. Methyl Thiazolyl Tetrazolium (MTT) and transwell assays were conducted to assess the cell viability and invasive abilities. Western blot was conducted to assess the protein levels of epithelial-mesenchymal transition (EMT) markers. Luciferase reporter assay was carried out to verify miR-150 directly binding to HMGA2 in SCC25 cells. RESULTS: MiR-150 was low expressed and HMGA2 was highly expressed in OSCC tissues and cells. Downregulation of miR-150 or upregulation of HMGA2 predicted poor prognosis of OSCC patients. MiR-150 overexpression inhibited the abilities of viability, invasive and the EMT by targeting HMGA2 in OSCC cells. HMGA2 was a target gene of miR-150 and its expression was regulated by altering the expression of miR-150 in OSCC cells. HMGA2 reversed partial roles of miR-150 on cell viability and invasion in OSCC. CONCLUSIONS: MiR-150 impaired cell viability, invasion and EMT via binding to HMGA2 of OSCC. Our research demonstrates that miR-150 plays a critical role in the progression of OSCC. miR-150 might be a candidate molecular marker and a novel therapy target for OSCC patients.

Effect of miR-132 on lung injury in sepsis rats via regulating Sirt1 expression.

OBJECTIVE: The aim of this study was to explore the effects of the expressions of micro ribonucleic acid (miR)-132 and sirtuin1 (Sirt1) on lung injury in sepsis rats, and to elucidate the regulatory relation between miR-132 and Sirt1. MATERIALS AND METHODS: The model of sepsis-induced lung injury was successfully established in rats via injection of lipopolysaccharide (LPS) into the caudal vein (model group). Before modeling, the rats were infused with miR-132 antagomir via the trachea (miR-132 antagomir group) or intraperitoneally injected with the Sirt1 activator (SRT1720) (SRT1720 group). Meanwhile, the rats injected with an equal volume of normal saline via the caudal vein were enrolled in the control group. The expressions of the inflammatory factors interleukin-6 (IL-6), IL-1ß, and tumor necrosis factor-α (TNF-α) were determined using enzyme-linked immunosorbent assay (ELISA). Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) and Western blotting were applied to detect gene and protein expressions in lung tissues, respectively. Targeted relationship between miR-132 and Sirt1 was explored using Luciferase reporter assay. In addition, tissue sections of the right lung were stained with hematoxylin-eosin (HE) to observe the degree of lung injury. RESULTS: The model of sepsis-induced lung injury was successfully established in rats by LPS. The results showed that the expressions of IL-6, IL-1ß, TNF-α and miR-132 rose significantly in lung tissues (p<0.01), whereas the expression of Sirt1 significantly declined (p<0.01). Lung injury was alleviated by miR-132 antagomir and SRT1720. Both miR-132 antagomir and SRT1720 significantly reduced the expressions of miR-132, IL-6, IL-1ß and TNF-α (p<0.01). However, the expression of Sirt1 was remarkably upregulated in rats with lung injury (p<0.01). Luciferase reporter gene assay indicated that miR-132 regulated Sirt1 in a targeted manner. CONCLUSIONS: MiR-132 may cause lung injury in sepsis rats by regulating the expression of Sirt1.

[Application of auditory brainstem response in hearing impairment and prognosis of neonatal hypoxic ischemic encephalopathy].

Objective:To investigate the effect of auditory brainstem response in hearing impairment and prognosis of neonatal hypoxic ischemic encephalopathy（HIE）. Method:Forty-five children with HIE admitted were enrolled. All patients were tested for auditory brainstem response and followed up for 3 years to observe the prognosis of the children. Result:A total of 90 ears were included in 45 children, of which 77 ears had hearing impairment, accounting for 85.6%; 38 ears had mild hearing impairment, accounting for 49.4%; 19 ears had moderate hearing impairment, accounting for 24.7%; 20 ears had severe hearing impairment, accounting for 26.0%; the difference was statistically significant（P<0.05）. Children with moderate and severe HIE had better hearing impairment than children with mild HIE. Serious, the difference was statistically significant（χ²=7.921, P=0.009）. A total of 90 ears were included in 45 patients, 40 of whom completed 3 years of follow-up. The prognosis of children with hearing threshold ≤60 dBnHL was worse than the threshold ≥61 dBnHL, and the difference was statistically significant（P=0.001）. Abnormal manifestations of auditory brainstem response were mainly in 13 children with mild HIE（76.5%） with poor differentiation or prolongation of Ⅰwave PL; 26 patients with moderate and severe HIE（92.9%） With Ⅲ, Ⅴ wave PL extension, Ⅲ-Ⅴ wave IPL extension, Ⅰ-Ⅲ wave IPL extension, Ⅰ-Ⅴ wave PL extension, Ⅰ-Ⅲ/Ⅲ-Ⅴ wave IPL wave interval ratio less than 1, and Ⅴ/Ⅰ amplitude less than 0.5. Conclusion:The auditory brainstem response in the hearing impairment and prognosis of HIE can effectively reflect the relationship between the severity of HIE and the severity of hearing impairment, and the relationship between the prognosis of children and the severity of hearing impairment. It can be used as an effective means to help determine the degree of HIE hearing loss and prognosis.

Impact of abnormal nutrition during pregnancy on the offspring hormone resistance.

OBJECTIVE: To investigate the impact of abnormal nutrition during pregnancy on the insulin and leptin resistance of adult offsprings. METHODS: The model of abnormal nutrition during pregnancy was established, and these rats were fed whole-course low-protein or high-nutrition. After natural childbirth, the birth weight of each newborn rat was measured. According to the determining birth weights, the newborn rats were assigned into the small for gestational age (SGA) and large for gestational age (LGA) groups as well as the healthy control group, respectively. There was a total of 36 randomly selected rats in each group. The levels of insulin and leptin and the insulin sensitivity index (ISI) were determined by enzymelinked immunosorbent assay 4 and 12 weeks post birth, respectively. RESULTS: In the low-protein group, the birth weight was significantly lower than in the control group (p<0.01) and 68.97% of the newborn rats were SGA; in the high-energy group, the birth weight of the newborn rats was significantly larger than in the control group (p<0.01), and 37.98% of the newborn were LGA. The body weights (BW) of the SGA 4 weeks post birth had no significant difference from that of the controls, while the perirenal fat weight (FW) and the FW/BW ratio were significantly larger than those of the controls (p<0.01 and p<0.05, respectively); however, the FW/BW of the LGA had no significant difference from that of the controls. Twelve weeks after birth, the BW of both SGA and LGA rats increased significantly compared to the controls (p<0.05 and p<0.01, respectively), and the FW/BW ratios of both were significantly larger than that of the controls (p<0.01). For the SGA rats 4 weeks post birth, the insulin and leptin level increased significantly (both p<0.05), while the ISI decreased significantly (p<0.05), with the occurrence of insulin resistance. For both SGA and LGA 12 weeks post birth, the insulin and leptin level significantly increased (both p<0.01). CONCLUSION: Abnormal nutrition during pregnancy could lead to abnormal birth weight, and both low and high birth weight could cause abdominal obesity as well as insulin and leptin resistance in adulthood, although through different mechanisms.

Expressions of HIF-1α and KISS-1 in patients with liver cancer and correlation analysis.

OBJECTIVE: To study the expressions of hypoxia-inducible factor-1α (HIF-1α) and tumor metastasis suppressor gene (KISS-1) in patients with liver cancer and to analyze the correlation between HIF-1α and KISS-1 and liver cancer. PATIENTS AND METHODS: 20 normal liver tissues and 30 liver cancer tissues in our hospital were selected. The expressions of HIF-1α and KISS-1 in normal liver tissues and liver cancer tissues were detected via immunofluorescence assay. The mRNA expressions of HIF-1α and KISS-1 in normal liver tissues and liver cancer tissues were detected via reverse transcription polymerase chain reaction (RT-PCR). The protein expressions of HIF-1α and KISS-1 in normal liver tissues and liver cancer tissues were detected via Western blotting. Differences of HIF-1α and KISS-1 expressions in normal liver tissues and liver cancer tissues were analyzed using SPSS 17.0 statistical software. RESULTS: Immunofluorescence assay, RT-PCR, and Western blotting, showed that HIF-1α was highly expressed in liver cancer tissues, and its expression level was significantly higher than that in normal liver tissues. However, the expression of KISS-1 in normal liver tissues was significantly higher than that in liver cancer tissues. The results of analysis of variance showed that the differences of HIF-1α and KISS-1 expressions in normal liver tissues and liver cancer tissues were statistically significant (p<0.01). CONCLUSIONS: The abnormal expressions of HIF-1α and KISS-1 are closely related to the development and progression of liver cancer, indicating that HIF-1α and KISS-1 have important research values in liver cancer, and the expressions of HIF-1α and KISS-1 can be used as the index of deterioration degree of liver cancer, providing a new clinical basis for diagnosis and treatment.

Characterization of the role of microRNA-517a expression in low birth weight infants.

The purpose of this study was to analyze the expression of the placenta-specific microRNA miR-517a in maternal serum and in placental tissue from low birth weight newborns and try to detect the effects of miR-517a expression on invasion potential of trophoblasts. Placental tissue and maternal serum were collected from both low birth weight newborns (n = 10) and normal birth weight newborns (n = 20). Expression of miR-517a was assessed in placenta and serum samples by real-time qRT-PCR. In addition, human trophoblast HTR8/SVneo cells were transfected with a miR-517a 2'-O-methyl oligonucleotide or a negative control RNA, and invasion was measured using transwell migration assays. Expression of miR-517a was significantly increased in placentas from low birth weight newborns (61.79 ± 23.06) in comparison with those of normal birth weight newborns (5.01 ± 1.97; P < 0.05). The expression of miR-517a was also increased in maternal serum isolated from the low birth weight newborn (25.78 ± 8.69) compared with the normal birth weight newborn (3.21 ± 1.07; P < 0.05). Overexpression of miR-517a significantly inhibited invasion of HTR8/SVneo cells (P < 0.05). These data indicate that miR-517a overexpression could potentially lead to low birth weight, likely through the inhibition of trophoblast invasion.