Identification, expression and DNA variation analysis of high affinity nitrate transporter NRT2/3 gene family in Sorghum bicolor / 生物工程学报

Nitrate is the main form of inorganic nitrogen that crop absorbs, and nitrate transporter 2 (NRT2) is a high affinity transporter using nitrate as a specific substrate. When the available nitrate is limited, the high affinity transport systems are activated and play an important role in the process of nitrate absorption and transport. Most NRT2 cannot transport nitrates alone and require the assistance of a helper protein belonging to nitrate assimilation related family (NAR2) to complete the absorption or transport of nitrates. Crop nitrogen utilization efficiency is affected by environmental conditions, and there are differences between varieties, so it is of great significance to develop varieties with high nitrogen utilization efficiency. Sorghum bicolor has high stress tolerance and is more efficient in soil nitrogen uptake and utilization. The S. bicolor genome database was scanned to systematically analyze the gene structure, chromosomal localization, physicochemical properties, secondary structure and transmembrane domain, signal peptide and subcellular localization, promoter region cis-acting elements, phylogenetic evolution, single nucleotide polymorphism (SNP) recognition and annotation, and selection pressure of the gene family members. Through bioinformatics analysis, 5 NRT2 gene members (designated as SbNRT2-1a, SbNRT2-1b, SbNRT2-2, SbNRT2-3, and SbNRT2-4) and 2 NAR2 gene members (designated as SbNRT3-1 and SbNRT3-2) were identified, the number of which was less than that of foxtail millet. SbNRT2/3 were distributed on 3 chromosomes, and could be divided into four subfamilies. The genetic structure of the same subfamilies was highly similar. The average value of SbNRT2/3 hydrophilicity was positive, indicating that they were all hydrophobic proteins, whereas α-helix and random coil accounted for more than 70% of the total secondary structure. Subcellular localization occurred on plasma membrane, where SbNRT2 proteins did not contain signal peptides, but SbNRT3 proteins contained signal peptides. Further analysis revealed that the number of transmembrane domains of the SbNRT2s family members was greater than 10, while that of the SbNRT3s were 2. There was a close collinearity between NRT2/3s of S. bicolor and Zea mays. Protein domains analysis showed the presence of MFS_1 and NAR2 protein domains, which supported executing high affinity nitrate transport. Phylogenetic tree analysis showed that SbNRT2/3 were more closely related to those of Z. mays and Setaria italic. Analysis of gene promoter cis-acting elements indicated that the promoter region of SbNRT2/3 had several plant hormones and stress response elements, which might respond to growth and environmental cues. Gene expression heat map showed that SbNRT2-3 and SbNRT3-1 were induced by nitrate in the root and stem, respectively, and SbNRT2-4 and SbNRT2-3 were induced by low nitrogen in the root and stem. Non-synonymous SNP variants were found in SbNRT2-4 and SbNRT2-1a. Selection pressure analysis showed that the SbNRT2/3 were subject to purification and selection during evolution. The expression of SbNRT2/3 gene and the effect of aphid infection were consistent with the expression analysis results of genes in different tissues, and SbNRT2-1b and SbNRT3-1 were significantly expressed in the roots of aphid lines 5-27sug, and the expression levels of SbNRT2-3, SbNRT2-4 and SbNRT3-2 were significantly reduced in sorghum aphid infested leaves. Overall, genome-wide identification, expression and DNA variation analysis of NRT2/3 gene family of Sorghum bicolor provided a basis for elucidating the high efficiency of sorghum in nitrogen utilization.

Effect of PM2.5 on inflammatory factors and pathology of nasal mucosa in a rat model of allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

Objective@#To investigate the effect of PM2.5 exposure on nasal inflammatory cytokines and nasal mucosal pathology in a rat model of allergic rhinitis (AR).@*Methods@#Twenty-four healthy female SD rats were randomly divided into 3 groups by random number table method, with 8 rats in each group: normal control group (NC group), ovalbumin (OVA) induced AR model (AR group), and AR model group inhaled to PM2.5 at 200 μg/m3, 3 h/d, for 30 d (ARE group). Nasal symptoms including sneezing, nasal rubs and nasal secretion were recorded. Levels of OVA specific IgE in serum, interleukin 6 (IL-6) and tumor necrosis factor-ɑ (TNF-ɑ) in nasal irrigating solution were measured by enzyme-linked immunosorbent assay (ELISA). The histopathological changes of nasal mucosa were observed by HE staining. SPSS 17.0 software was used to analyze the data.@*Results@#The number of sneezing, nasal rubs and the amount of nasal secretion in the ARE group were significantly higher than that in the AR group and the NC group (number of sneezing (15.38±1.68) times/15 min vs (11.63±1.13) times/15 min vs (1.75±0.71) times/15 min, number of nasal rubs (27.75±2.12) times/15 min vs (21.25±2.96) times/15 min vs (5.25±1.04) times/15 min, amount of nasal secretion (18.90±2.07) mg vs (13.83±1.81) mg vs (3.78±0.41) mg, F values was 236.089, 224.139, 183.971, respectively, all P<0.001). Statistically significant differences in OVA specific IgE, IL-6 and TNF-ɑ levels were observed in ARE group exceeded AR group and NC group (OVA specific IgE (25.42±2.51) ng/ml vs (18.07±1.07) ng/ml vs (1.47±0.26) ng/ml, IL-6 (123.30±18.86) pg/ml vs (63.49±11.29) pg/ml vs (16.87±3.29) pg/ml, TNF-ɑ (162.50±38.15) pg/ml vs (72.96±11.28) pg/ml vs (27.52±4.15) pg/ml, F values was 481.604, 138.277, 63.938, respectively, all P<0.001). HE staining showed that the nasal epithelial cells of NC group were intact and neatly arranged. Nasal mucosa epithelial cells were arranged in disorder in AR group, with tissue structure swelling. Partial shedding of nasal epithelial cells, mucosal basement membrane thickening, submucosal tissue interstitial edema, vasodilation and gland hyperplasia were found in ARE group.@*Conclusion@#An increase inflammatory factors level such as IL-6 and TNF-ɑ aggravates pathological damage of nasal mucosa in a rat model of AR by exposure to PM2.5.

The role of autophagy in PM2.5-induced inflammation in human nasal epithelial cells / 中华耳鼻咽喉头颈外科杂志

Objective@#To explore the role of autophagy in PM2.5-induced inflammation in human nasal epithelial cells and related mechanism.@*Methods@#Human nasal epithelial cells were exposed to different concentration of PM2.5 for different times, and the expression levels of microtubule-associated protein-1 light chain-3 Ⅱ (LC3 Ⅱ) and Beclin1 proteins were measured by Western blot. The typical autophagosome and autolysosome were observed by using transmission electron microscopy (TEM). To observe autophagic flux, mRFP-GFP-LC3 plasmid was transfected to nasal epithelial cells and the punctate staining of mRFP-GFP-LC3 were determined by confocal laser scanning microscope. The expression of inflammatory cytokines interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) in cell culture supernatant were assessed by enzyme-linked immunosorbent assay (ELISA). To assess the role of autophagy in PM2.5-mediated inflammation, autophagy related gene Atg5 and Beclin-1 were silenced by siRNA knockdown, and inflammatory cytokines were analyzed.GraphPad Prism 6.0 was used for statistical analysis.@*Results@#PM2.5 exposure increased the expression of LC3 Ⅱ and Beclin-1 proteins in a dose- (in PM2.5 group with concentration of 0, 15, 30, 60, 120 μg/ml, the expression of LC3 Ⅱ was 0.021±0.001(±s), 0.037±0.002, 0.058±0.005, 0.075±0.006, 0.085±0.004, respectively, F=126.8, P<0.05; the expression of Beclin-1 was 0.002±0.000, 0.003±0.000, 0.005±0.000, 0.007±0.001, 0.008±0.001, respectively, F=137.3, P<0.05) and time-dependent manner (in PM2.5 group with exposure time of 0, 3, 6, 12, 24 h, the expression of LC3Ⅱ was 0.160±0.007, 0.222±0.003, 0.251±0.015, 0.483±0.029, 0.585±0.035, respectively, F=215.3, P<0.05; the expression of Beclin-1 was 0.059±0.002, 0.080±0.002, 0.087±0.002, 0.183±0.007, 0.228±0.005, respectively, F=137.3, P<0.05) in human nasal epithelial cells. TEM analysis showed typical autophagosome and autolysosome in cells after PM2.5 exposure for 24 h. PM2.5 significantly increased the number of yellow and red dots representing autophagosomes and autolysosomes respectively, indicating autophagic flux was elevated. Moreover, PM2.5 enhanced the secretion of inflammatory cytokines such as IL-6 and TNF-α, which was dramatically prevented by Atg5-siRNA and Beclin-1-siRNA.@*Conclusion@#Autophagy plays an important role in PM2.5-caused inflammation response in nasal epithelial cells, which can induce release of inflammatory factors such as IL-6 and TNF-α and advance the inflammatory reaction.

Chinese Society of Allergy Guidelines for Diagnosis and Treatment of Allergic Rhinitis

Allergic rhinitis (AR) is a global health problem that causes major illnesses and disabilities worldwide. Epidemiologic studies have demonstrated that the prevalence of AR has increased progressively over the last few decades in more developed countries and currently affects up to 40% of the population worldwide. Likewise, a rising trend of AR has also been observed over the last 2–3 decades in developing countries including China, with the prevalence of AR varying widely in these countries. A survey of self-reported AR over a 6-year period in the general Chinese adult population reported that the standardized prevalence of adult AR increased from 11.1% in 2005 to 17.6% in 2011. An increasing number of original articles and imporclinical trials on the epidemiology, pathophysiologic mechanisms, diagnosis, management and comorbidities of AR in Chinese subjects have been published in international peer-reviewed journals over the past 2 decades, and substantially added to our understanding of this disease as a global problem. Although guidelines for the diagnosis and treatment of AR in Chinese subjects have also been published, they have not been translated into English and therefore not generally accessible for reference to non-Chinese speaking international medical communities. Moreover, methods for the diagnosis and treatment of AR in China have not been standardized entirely and some patients are still treated according to regional preferences. Thus, the present guidelines have been developed by the Chinese Society of Allergy to be accessible to both national and international medical communities involved in the management of AR patients. These guidelines have been prepared in line with existing international guidelines to provide evidence-based recommendations for the diagnosis and management of AR in China.

Study on apoptosis of breast cancer cells induced by green tea / 中国基层医药

Objective To study the apoptosis of breast cancer cells induced by green tea and the preventive effect of green tea on cancer.Methods Catecholamine,the main components of green tea was added into human breast cancer cell line(MDA-MB-231) with different concentrations,and then human breast cancer cell line was measured by MTT assay,comet assay,flow cytometry and caspase-3 activity assay respectively.Results After treatment with 0.2mmol/L EGCG for 48 h,the cell proliferation was inhibited in the experimental group in MTT assay[minimum absorbance value (0.391±0.041),t=4.223 P<0.01].In comet assay,cells treated with 0.2mmol/L EGCG for 48 h in the experimental group showed a fairly long tail[control group average value (4.92±0.64)μm,the experimental group average value (18.76±1.37)μm,P=0.003].The rate of cell apoptosis increased significantly by testing MDA-MB-231 exposed to EGCG with flow cytometry.The apoptotic rate of the cells exposed to 0.2mmol/L EGCG for 48 h in the experimental group was (29.370±1.485)(t=11.125,P<0.01).EGCG induced apoptosis and caspase-3 activity rate was dependent on time and dose.The OD value of caspase-3 observed by the colorimetric method in cells exposed to 0.2mmol/L EGCG for 48 h in the experimental group was (0.144±0.045)(t=5.321,P<0.01).Conclusion EGCG may play a role in the treatment and prevention of breast cancer by affecting apoptosis.

Application of thymalfasin in non-operative treatment of cervical cancer / 中国基层医药

Objective To observe the effect of thymalfasin on non-operative treatment of cervical cancer.Methods Seventy-eight patients with advanced cervical cancer were divided into two groups according to the digital table,each group in 39 cases.The two groups were treated with concurrent chemoradiotherapy.The observation group was given 1.6 mg of thymalfasin subcutaneously each Monday to Friday.While the control group received no additional treatment.And then the two groups were compared in terms of curative effects,adverse reactions,immune states and cellular immune functions.Results The results showed that the differences of curative effects[there were 23 patients in the control group,as compared with 31 patients in the observation group,achieved a complete response(CR),x2=3.852,P<0.05],adverse reactions(grade Ⅲ or Ⅳ radiation-induced enteritis occurred in 4 patients in the observation group,and 12 patients in the control group,x2=5.032,P<0.05;Grade Ⅲ or Ⅳ leukopenia occurred in 16 patients in the observation group,and 25 patients in the control group,x2=4.165,P<0.05;Grade Ⅲ or Ⅳ nausea,vomit occurred in 6 patients in the observation group,and 14 patients in the control group,x2=4.303,P<0.05),immune states and cellular immune functions between the two groups were significant[the counts of CD+3(t=9.236,P<0.05),CD+4(t=7.445,P<0.05),CD+4/CD+8(t=7.445,P<0.05) and NK(t=9.256,P<0.05)]were significantly higher in the observation group after treatment.Conclusion In the treatment of advanced cervical cancer,concurrent chemoradiotherapy combined with thymalfasin can improve the curative effect,reduce the side effects,improve the quality of life and enhance the immunity.

Study on inhibition of genistein on doxorubicin-resistant bladder tumor cells T24 / 肿瘤研究与临床

Objective To establish multi-drug resistant bladder (MDR) tumor T24 cell lines and to assess their resistant characteristics.To observe effect of genistein on doxorubicin (ADM) resistant cell lines T24/ADM.Methods Bladder tumor T24 cell line was exposed to ADM in the culture medium for the establishment of drug resistant cell lines:concentrations of ADM was stepwise increased for long exposure.Morphologic studies were performed with optical microscopy.Drug sensitivities were determined by MTT.Results Six months were taken to establish drug resistant cell lines T24/ADM.No obvious morphologic changes were observed between resistant and parental cell. But drug resistances to ADM, 5-Fu,cyclophosphamide and cisplatin were increased,and resistance index were 15.79,4.68,5.53 and 3.81,respectively.Among all groups,there were significant differences.After genistein was used to T24/ADM cells,the IC50 value of genistein was 40 μg/ml.The proliferation cells were induced by genistein at the concentration of 20-100 μg/ml. Conclusion Genistein can inhibit human urinary bladder cancer T24/ADM cell proliferation at some concentration.

Evaluation of Surgical Approaches for Benign Ovarian Cyst / 中国医科大学学报

Objective To evaluate the clinical value of laparotomy,transvaginal surgery,and lapaioscopic surgery in treating benign ovarian cyst.Methods We retrospectively analyzed 227 patients with benign ovarian cyst who underwent laparotomy,transvaginal surgery,or laparoscopic surgery.The duration of the operation,amount of blood loss,hospitalization time,anal exhaust time,temperature recovery time.outof-bed time,usage of analgesics,postoperative morbidity,and hospitalization cost were analyzed.Results There were significant differences in above indicators between the patients undergoing transvaginal surgery or laparoscopic surgery and those undergoing laparotomy.Conclusion Transvaginal surgery and laparoscopic surgery are minimally invasive and safe in treating benign ovarian cyst.These 3 surgical approaches have distinct advantages and disadvantages.The clinicians should choose the appropriate surgical approach for benign ovarian cyst according to the patient's condition.

Endcapping of Octadecyl Bonded Silica by Atomic Layer Deposition for Separation of Basic Compounds / 分析化学

Atomic layer deposition has been employed for postsynthesis treatment of octadecyl bonded silica (ODS) as a typical reversed-phase stationary phase for high performance liquid chromatography (HPLC) ,in an effort to alleviate peak tailing for basic compounds. With hexamethyldisilazane( HMDS) as an endcapping agent,the ODS packing materials were heated to 250℃,and maintained at that temperature for 6 h,affording packing materials which are highly inert to basic compounds. Chromatographic performance in terms of retention factor and tailing factor of the resulting phases was evaluated using phenol/pyridine and naphthalene/ami-triptyline pairs as probes. The results were compared with those for the same packing treated by liquid phase method and for the commercial products including Zorbax SB-C_(18) and Kromasil C_(18). The separation characteristics of the ODS phase obtained by atomic layer deposition appear to be superior to that by liquid phase method and,comparable with or even better than the commercial products studied. Being solvent-free process amenable to mass production,the described method provides an economical and eco-friendly approach to manufacturing HPLC packing materials on industrial scale.

Analysis of 30 Cases of Continuous Positive Airway Pressure Curing Overlap Syndrome / 医学研究杂志

Objective To study the validity of continuous positive airway pressure(CPAP)curing overlap syndrome.Methods Comparisons were made in the respiratory disturbance index(RDI)the longest apnea time the longest hypopnea time etc between before and after-treatment.Results The respiratory disturbance index(RDI)、the longest apnea time、the longest hypopnea time were signifi- candy different(P＜0.05).Conclusion Continuous positive airway pressure is an effective method to treat overlap syndrome.

An Etiological Study on Hearing Loss of 813 Deaf Students in Hubei Province / 听力学及言语疾病杂志

Objective In order to study the etiology and the hearing status of the deaf students in Hubei province, a survey was carried out from April 1999 to June 2000.Methods A total number of 813 deaf students in Hubei province were examined with audiometer and investigated through questionnaire. The pedigress analysis was conducted in deaf students with family history.Results The pedigrees of 227 familes with deafness were obtained, the inheritance pattern of 167 families could be ascertained. 232(28.5%) deaf students were diagnosed congenital deafness, 581 (71.5%) students were diagnosed acquired deafness. The degrees of deafness could be ascertained with 359(44.1%) students of profound deafness, 323(39.7%) students of severe deafness, 111(13.7%) students with moderate to severe deafness, 11(1.4%) students of moderate deafness, and 9(1.1%) students of mild deafness.Conclusion The hearing loss of deaf students is very serious, and genetic factor and ototoxic antibiotics were a principal causation in the occurrence of deafness.