1.Study on expression and clinical significance of hypoxia-inducible factor 1α and glucose transporter-1 and their relationship with the clinicopathological characteristics in elderly patients with primary lung cancer
Ling WANG ; Chunlai FENG ; Ningzheng DONG
Chinese Journal of Geriatrics 2009;28(2):128-132
Objective To explore the expression and clinical significance of hypoxia-inducible factor 1α(HIF-1α)and glucose transporter-1(GLUT-1)in human lung adenocarcinoma cell line A549 and in elderly primary lung cancer tissue under hypoxia. Methods MTT assay was used to test the effect of CoCl2-induced hypoxia on the growth activity of lung adenocarcinoma cell line A549.Reverse transcription-polymerase chain reaction analysis(RT-PCR)and Western blot were used to determine the changes of HIF-1α and GLUT-1 in lung adenocarcinoma cell line A549 under hypoxia.The expressions of HIF-1α and GLUT-1 were detected by using immunohistochemical method and RT-PCR in 60 specimens of lung carcinoma and 26 benign lung tissues in elderly patients with pulmonary diseases.Their interrelations and their relationship with clinicopathologicaI characteristics of the tumors were analyzed. Results The growth activity of lung adenocarcinoma cell line A549 was increased under hypoxia.RT-PCR analysis indicated that the mRNA expressions of HIF-1α and GLUT-1 were 0.69±0.08,0.65±0.09 after hypoxia exposure for 24 hours,and 0.99±0.16,0.83±0.11 after hypoxia exposure for 48 hours,which were higher than those in control group (0.47±0.11,0.40±0.08,P<0.01).The results from Western blot showed that the protein expressions of HIF-1α and GLUT-1 were 1.57±0.29,1.66±0.28 after hypoxia exposure for 24 hours,and 2.31±0.46,2.22±0.34 after hypoxia exposure for 48 hours,which were higher than those in control group(0.93±0.07,1.06±0.17,P<0.01).In specimens of lung carcinoma,the positive expression rates of mRNA and proteins of HIF-1α and GLUT-1 were higher than those in benign lung tissues(all P<0.01).Compared with the mRNA and protein levels of HIF-1a and GLUT-1 in lung carcinoma tissues of Ⅰ and Ⅱ stage,there were marked increases in Ⅲ and Ⅳ stage (all P<0.05).The expressions of HIF-1α and GLUT-1 in lymph node metastasis group were higher than those in group without lymph node metastasis(all P<0.05).There were positive correlations between the mRNA and protein expressions of HIF-1α and between the mRNA and protein expressions of GLUT-1(r1=0.527,r2=0.408,respectively,both P<0.01). Conclusions CoCl2 can up-regulate the expressions of HIF-1α and GLUT-1 in lung adenocarcinoma cell line A549 by inducing hypoxia.The hypoxia-mediated effects on them have the potential to make lung cancer cells further grow and promote hypoxie tolerance.The overexpressions of HIF-1α and GLUT-1 are closely correlated with the progression of lung carcinoma.Combined detection of HIF-1α and GLUT-1 may be helpful for the diagnosis and prognosis prediction of primary lung cancer in elderly patients.
2.Pedicled muscular flap for treatment of pyothorax-resulted wound.
Yong ZHANG ; Zihao FENG ; Yanwen YANG ; Chunlai LU ; Di GE ; Fazhi QI
Chinese Journal of Plastic Surgery 2014;30(6):428-431
OBJECTIVETo evaluate the therapeutic effect of pedicled muscular flaps combined with partial thoracotomy for treatment of pythorax-resulted wounds and defects.
METHODS35 cases with pythorax-resulted refractory infected wound and dead space, were retrospectively analyzed. The wound and dead space were eliminated with pedicled muscular flaps, including latissimus dorsi muscular flaps and serratus anterior muscle flaps in 21 patients, pectoralis major muscle flaps with rectus muscle flaps in 8 patients, latissimus dorsi muscul flaps and vertical rectus muscle flaps in other 6 patients.
RESULTS34 flaps survived completely except for 1 verticle rectus muscular flap with partial necrosis at distal end. Primary healing was achieved in 29 cases. The other 6 cases were discharged with drainage. Among the 6 cases, 3 cases healed spontaneously, 2 cases underwent reoperation, 1 case need long-term drainage in the fistula.
CONCLUSIONPedicled muscular flap, combined with partial thoracotomy can effectively treat refractory wound and dead space resulted by pythorax.
Combined Modality Therapy ; methods ; Empyema, Pleural ; surgery ; Humans ; Myocutaneous Flap ; transplantation ; Pectoralis Muscles ; transplantation ; Reoperation ; Retrospective Studies ; Superficial Back Muscles ; transplantation ; Thoracotomy ; methods ; Wound Healing
3.Sequence analysis and genotyping of genital Chlamydia trachomatis among patients with suspected-Neisseria gonorrhoeae
Juanjuan ZHANG ; Guanglu ZHAO ; Lijun ZHANG ; Feng WANG ; Fuchang HONG ; Lina LAN ; Xiaobing WU ; Xiaohua TAO ; Chunlai ZHANG ; Ciyong LU ; Tiejian FENG
Chinese Journal of Microbiology and Immunology 2010;30(12):1159-1163
Objective To understand the prevalence rate of genital Chlamydia trachomatis among a population with suspected-Neisseria gonorrhoeae infection,the distribution of Chlamydia trachomatis genotypes,assess changes in omp1 sequences among patients with Neisseria gonorrhoeae and Chlamydia trachomatis coinfections.Methods Four hundred and one swabs were collected.Chlamydia trachomatis and Neisseria gonorrhoeae were detected by Roche Amplicor System.DNA were extracted from those samples and were amplified by nested PCR.PCR products were sequencing and analyzed by software Mega4.0.Results The prevalence of genital Chlamydia trachomatis infection,Neisseria gonorrhoeae infection and coinfection with genital gonorrhoea and genital chlamydia were 82.3%,24.2% and 21.7% each.Eight genotypes were identified in 73 sequences,including E(27.4%),G/Ga(23.3%),D/Da(16.4%),F(13.7%),J (11.0%),H(5.5%),B and K(each 1.4%).Sequencing analysis showed that 3 cases(4.1%) had missense mutation,including genotype D/Da,E,G/Ga.Genotypes F,H,J and K were more variable,however,most of them were silent mutation.Conclusion The prevalence rate of genital Chlamydia trachomatis among a population with Neisseria gonorrhoeae infection was high.The most common genotypes were genotype E,G/Ga,D/Da and F; Sequencing analysis has provided a tool for the molecular epidemiology of genital Chlamydia trachomatis infections.
4.Contact tracing of pregnant women infected with syphilis and the associated factors.
Xiaobing WU ; Fuchang HONG ; Email: HFC0755@SINA.COM. ; Chunlai ZHANG ; Tiejian FENG ; Lina LAN ; Yingzhou YANG
Chinese Journal of Preventive Medicine 2015;49(12):1067-1072
OBJECTIVETo investigate the current status of contact tracing among pregnant women infected with syphilis and to analyze the associated factors from patients' perspective.
METHODSPregnant women who aged 18 years old, receiving prenatal care services in Shenzhen, diagnosed with syphilis according to national diagnostic criteria (WS 273-2007) from 2008 to 2011 were recruited and the total number was 3 551. Information of both pregnant women (including demographic information, laboratory results, syphilis diagnosis, and personal life history) and their partners (including results of partner tracing and laboratory examination) were collected with structured questionnaire by face-to-face interview. Multivariate non-conditional logistic regression model was applied to analyze the factors associated with partners' contract tracing. Odds ratio (OR) and its 95% confidential interval (95%CI) were calculated.
RESULTSThe average age was 28.72 among recruited 3 551 syphilis-infected pregnant women, with standard deviation of 5.21 and range of 18 to 40. Totally 2 550 partners attended antenatal clinics and received syphilis examination, with a contact tracing rate of 71.81%. The OR(95%CI) was 1.70(1.26-2.30) for pregnant women with college or above education when comparing with those with senior high school or below education. The OR(95%CI) was 0.57(0.45-0.71) for those unmarried or divorce when comparing with those married. The OR (95% CI) was 0.73(0.56-0.94) and 0.65(0.53-0.81) for those screened in 28-36 gestational weeks and those screened in >36 gestational weeks or right before delivery separately, when comparing with those screened in ≤27 gestational weeks. The OR (95% CI) was 1.45(1.14-1.84) for those having received sufficient treatment before pregnancy when comparing with those diagnosed with latent syphilis. The OR(95%CI) was 0.31 (0.24-0.39) for those having no treatment or not standardized treatment when comparing with those having standardized penicillin treatment. The OR(95%CI) was 0.53 (0.43-0.65) for those terminating the pregnancy when comparing with those giving birth to a baby. The OR(95%CI) was 1.24 (1.05-1.46) for those having two or more love histories when comparing with those having only one love history. The OR(95%CI) was 4.74 (3.54-6.35) for those intending to disclose the serostatus to their partner when comparing with those showing unwillingness to disclose the serostatus.
CONCLUSIONThe successful rate of contact tracing among pregnant women infected with syphilis was high. The implementation of contact tracing may be affected by many factors, including patients' education levels, marital status, gestational weeks of screening, stage of syphilis, treatment status, outcome of pregnancy, number of love histories, and willingness of serostatus disclosure.
Ambulatory Care Facilities ; China ; epidemiology ; Contact Tracing ; Female ; Humans ; Logistic Models ; Odds Ratio ; Pregnancy ; Pregnancy Complications, Infectious ; epidemiology ; Prenatal Care ; Sexual Partners ; Syphilis ; epidemiology
5.Identification, expression and DNA variation analysis of high affinity nitrate transporter NRT2/3 gene family in Sorghum bicolor.
Shanshan ZHAO ; Zhiqiang GUO ; Lixun ZHU ; Jiali FAN ; Bohui YANG ; Wenting CHAI ; Huiqiong SUN ; Fan FENG ; Yuexiu LIANG ; Chunlei ZOU ; Xiaodong JIANG ; Weijun ZHAO ; Jinhui LÜ ; Chunlai ZHANG
Chinese Journal of Biotechnology 2023;39(7):2743-2761
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.
Nitrate Transporters
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Nitrates/metabolism*
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Sorghum/metabolism*
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Anion Transport Proteins/metabolism*
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Phylogeny
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Protein Sorting Signals/genetics*
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Nitrogen/metabolism*
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DNA
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Gene Expression Regulation, Plant
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Plant Proteins/metabolism*