Phylogenetic Analysis of Citrus tristeza virus Isolates of Wild Type Citrus in China

The genetic variation and phylogenetic relationships of Citrus tristeza virus （CTV） isolates collected from Chinese wild type citrus were analyzed by comparing the sequences of nine genomic regions （p23, p20, p13, p18, p25, p27, POL, HEL and k17） with the CTV isolates of cultivated citrus from different countries. The results showed that the divergence pattern of genomic RNA of the CTV isolates from wild type citrus was similar to that of other isolates from cultivated citrus, the 3′ proximal region was relatively conserved, and the 5′ proximal region had greater variability. The nine genomic regions of CTV isolates analyzed were found to have been under purifying selection in the evolution process. Phylogenetic analysis showed that the eleven Chinese wild CTV isolates were located at different clades and did not relfect their geographical origins, suggesting genetic diversity among the Chinese wild CTV populations. These results will aid in the understanding of molecular evolution of the Chinese CTV populations.

Changes in gene-expression profiles of colon carcinoma cells induced by wild type K-ras2

AIM: To further elucidate the possible molecular biological activity of wild type K-ras2 gene by detecting changes in wild type K-ras2 gene-induced gene-expression profiles of colon carcinoma cells using cDNA microarray techniques. METHODS: Total RNA was isolated from peripheral blood of health volunteers. Reverse transcription of RNA and polymerase chain reaction were used to synthesize wild type K-ras2 cDNA. K-ras2 cDNA fragment was cloned into a T easy vector and sequenced. A eukaryotic expression vector pCI-neo-K-ras2 was constructed and transfected to Caco2 cell line using the liposome method. Finally, mRNA was isolated, reverse-transcribed to cDNA from pCI-neo-K-ras2 or pCI-neo blank vector-transfected Caco cells, and analyzed by cDNA microarray assay. RESULTS: Restriction enzyme analysis and DNA sequencing verified that the constructed expression vector was accurate. High-quality RNA was extracted and reverse transcribed to cDNA for microarray assay. Among the 135 genes, the expression was up-regulated in 24 and down-regulated in 121. All these differentially expressed genes were related to cell proliferation, differentiation, apoptosis and signal transduction. CONCLUSION: Differentially expressed genes can be successfully screened from wild type K-ras2-transfected colon carcinoma cells using microarray techniques. The results of our study suggest that wild type K-ras2 is related to the negative regulation of cell proliferation, metabolism and transcriptional control, and provide new clues to the further elucidation of its possible biological activity.

Morphological Characteristics and Nutritional Values of Wild Types of Sago Mushrooms (<i>Volvariella</i>sp.) That Growth Naturally in Manokwari, West Papua

Sago mushrooms (SMs) are an edible fungus that is favorite food for community in Papua and West Papua, Indonesia in particular. This work aims to determine the morphological characteristics and nutritional value of SMs growing naturally in Manokwari. Morphological characteristic of SMs that grows wild in Manokwari has an average of pileus diameter 9.53 cm, pileus weight 21.53 grams, and pileus color. The pileus color is divided into three colors: outer circle color is RHS163D, middle circle color is RHS199B, and inner circle color is RHS199A. Average of stipe diameter, length, and weight is 1.00 cm, 10.43 cm, and 9.15 gram respectively. Stipe is a yellowish white color (RHS155B). SMs nutrient content that grows naturally in Manokwari is potassium 1394.02 milli-grams and calcium 13.37 milligrams per 1000 grams fresh weight. The others nutrient contents of SMs that are measured are fat 1.01 grams, protein 1.30 grams, carbohydrates 0.18 grams, phosphorus 0.34 milligrams, and energy 15.01 kilocalories per 100 grams of fresh weight.

Oral Vinorelbine as Switch Maintenance Therapy versus Best Supportive Care in Patients with Advanced Adenocarcinoma Non-Small Cell Lung Cancer EGFR Wild Type

Background: This study was performed to evaluate the efficacy and safety of switch maintenance therapy with oral vinorelbine in advanced non-small cell lung cancer (NSCLC) with adenocarcinoma limited to epidermal growth factor receptor (EGFR) wild type. Materials and Methods: In this single randomized trial, patients with advanced stage (IIIB and IV) NSCLC with adenocarcinoma EGFR wild-type status, treated with 6 cycles of platinum based chemotherapy. Patients did not show progression after first-line chemotherapy were randomly assigned to receive switch maintenance with vinorelbine (80 mg/m2, day 1, 8) (group I) or the best supportive care until disease progression (group II). Results: The median progression free survival (PFS) was 9.7 months for group I versus 5.7 months for group II with statistically significant difference between both groups [HR = 1.15;95% CI 1.19 to 1.49;P value = 0.002], while the median overall survival (OS) was 13.2 months for group I versus 11.9 months for group II with no statistically significant differences between both groups [HR = 1.24;95% CI 1.05 to 1.46;P value = 0. 3]. The patients who received oral vinorelbine had tolerable toxicity profile. Conclusion: Switch maintenance therapy with oral vinorelbine, though improve PFS, did not improve OS in patients with NSCLC with adenocarcinoma EGFR wild type.

KIT and platelet-derived growth factor receptor α wild-type gastrointestinal stromal tumor associated with neurofibromatosis type 1: Two case reports

BACKGROUND Gastrointestinal stromal tumors(GISTs) associated with neurofibromatosis are uncommon compared to their gastrointestinal counterparts. Patients with neurofibromatosis type 1(NF-1) have an increased risk of developing gastrointestinal tumors, including rare types such as GIST.CASE SUMMARY A 60-year-old male Chinese patient was diagnosed with NF-1 10 years ago and presented with upper abdominal discomfort and black stools. Endoscopic ultrasonography and an enhanced abdominal computed tomography scan revealed a mass located 4 cm from the muscular layer of the descending duodenum. A 59-year-old Chinese woman who was diagnosed with NF-1 25 years ago presented with sudden unconsciousness and black stools. Multiple masses in the duodenum were noted by echogastroscopy and an enhanced abdominal computed tomography scan. Both patients presented with cutaneous neurofibromas. The histologic examination of tumors from both patients revealed spindle cells and low mitotic activity. Immunohistochemically, the tumor cells showed strong positivity for KIT(CD117), DOG-1, CD34, and Dehydrogenase Complex Subunit B, and negativity for SMA, desmin, S-100, and β-catenin. None of the six tumors from two patients had KIT exon 9, 11, 13, or 17 or platelet-derived growth factor receptor α exon 12 or 18 mutation, which is a typical finding for sporadic GISTs. None of the six tumors from the two patients had a BRAFV600 E mutation. The patients were alive and well during the follow-up period(range:0.6-5 yr).CONCLUSION There have been only a few previous reports of GISTs associated with NF-1.Although GISTs associated with NF-1 have morphologic and immunohistochemical similarities with GISTs, the pathogenesis, incidence,genetic background, and prognosis are not completely known. A medical history of NF-1 in a patient who has gastrointestinal bleeding or anemia and an intraabdominal mass with nonspecific computed tomography features may help in diagnosing GIST by virtue of the well-known association of these two entities.Molecular genetic studies of cases indicated that GISTs in NF-1 patients have a different pathogenesis than sporadic GISTs.

Reversal of Multidrug Resistance and Inhibition of Phosphorylation of AKT in Human Ovarian Cancer Cell Line by Wild-type PTEN Gene

The reversing effect of wild-type PTEN gene on resistance of C 13K cells to cisplatin and its inhibitory effect on the phosphorylation of protein kinase B （AKT） were studied. The expression of PTEN mRNA and protein in OV2008 cells and C13K cells were semi-quantitatively detected by using RT-PCR and Western blotting. Recombinant eukaryotic expression plasmid containing human wild-type PTEN gene was transfected into C13K cells by lipofectamine2000. The expression of PTEN mRNA was monitored by RT-PCR and the expression of PTEN, Akt, p-Akt protein were ana- lyzed by Western blotting in PTEN-transfected and non-transfected C13K cells. Proliferation and chemosensitivity of cells to DDP were measured by MTT, and cell apoptosis was detected by flow cytometry after treatment with cisplatin. The expression of PTEN mRNA and protein in OV2008 cells were significantly higher than those in C13K cells. After transfection with PTEN gene for 48 h, the expression of PTEN mRNA and protein in C 13K cells were 2.04 ± 0.10, 0.94± 0.04 respectively and the expression of p-Akt protein （ 0.94± 0.07） was lower than those in control groups （1.68 ±0.14, 1.66± 0.10） （P〈 0.05）. The IC50 of DDP to C 13 K cells transfected with PTEN （7.2± 0.3 la mol/L） was obviously lower than those of empty-vector transfected cells and non-transfected cells （12.7±0.4 lamol/1, 13.0±0.3 lamol/L） （P〈0.05）. The apopototis ratio of wild-type PTEN-transfected, empty vector transfected and non-transfected C13K cells were （41.65___0.87）%, （18.61 ±0.70）% and （15.28±0.80）% respectively, and the difference was statistically significant （P〈0.05）. PTEN gene plays an important role in ovarian cancer multidrug resistance. Transfection of PTEN could increase the expression of PTEN and restore drug sensitivity to cisplatin in human ovarian cancer cell line C 13K with multidrug-resistance by decreasing the expression of p-Akt.

Construction and expression of eukaryotic plasmids containing lamivudine-resistant or wild-type strains of Hepatitis B Virus genotype C

AIM:To construct eukaryotic expression plasmids of full-length Hepatitis B Virus(HBV) genotype C genome,which contain lamivudine-resistant mutants(YIDD,YVDD) or wild-type strain(YMDD) ,and to observe the expression of HBV DNA and antigens [hepatitis B surface antigen(HBsAg) and hepatitis B e antigen(HBeAg) ] of the recombinant plasmids in HepG2 cells. METHODS:Three HBV full-length genomes were amplified from the plasmids pMD18T-HBV/YIDD,pMD18T-HBV/YVDD and pMD18T-HBV/YMDD,using PCR. Three recombinant plasmids were generated by inserting each of the PCR products into the eukaryotic expression vector pcDNA3.1(+) ,between the EcoRI and HindⅢ sites. After being characterized by restriction endonuclease digestion,and DNA sequence analysis,the recombinant plasmids were transfected into HepG2 cells. At 48 and 72 h post-transfection,the levels of intracellular viral DNA replication were detected by real-time PCR,and the expression of HBsAg and HBeAg in the cell culture supernatant was determined by ELISA. RESULTS:Restriction endonuclease digestion and DNA sequence analysis confirmed that the threerecombinant plasmids were correctly constructed. After transfecting the plasmids into HepG2 cells,high levels of intracellular viral DNA replication were observed,and HBsAg and HBeAg were secreted into the cell culture supernatant. CONCLUSION:Eukaryotic expression plasmids pcDNA3.1(+) -HBV/YIDD,pcDNA3.1(+) -HBV/YVDD or pcDNA3.1(+) -HBV/YMDD,which contained HBV genotype C full-length genome,were successfully constructed. After transfection into HepG2 cells,the recombinant plasmids efficiently expressed HBV DNA,HBsAg and HBeAg. Our results provide an experimental basis for the further study of HBV lamivudine-resistant mutants.

Stem cell factor-mediated wild-type KIT receptor activation is critical for gastrointestinal stromal tumor cell growth

AIM: To clarify the biological role of stem cell factor (SCF)-mediated wild-type KIT receptor activation in gastrointestinal stromal tumor (GIST) growth. METHODS: The co-expression of wild-type KIT receptor and SCF was evaluated in 51 GIST samples using mutation analysis and immunohistochemistry, and the results were correlated with clinicopathological param- eters, including the mitotic count, proliferative index (Ki-67 immunohistochemical staining), mitotic index (phospho-histone H3 immunohistochemical staining) and apoptotic index (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling). Using primary cultured GIST cells, the effect of SCF-mediated wild-type KIT receptor activation was determined by western blotting, methyl thiazolyl tetrazolium (MTT), and apoptosis assays. RESULTS: We found that wild-type KIT receptor and SCF protein were expressed in 100% and 76.5% of the 51 GIST samples, respectively, and the co-expression of wild-type KIT receptor and SCF was associated with known indicators of poor prognosis, including larger tumor size (P = 0.0118), higher mitotic count (P = 0.0058), higher proliferative index (P = 0.0012), higher mitotic index (P = 0.0282), lower apoptosis index (P = 0.0484), and increased National Institutes of Health risk level (P = 0.0012). We also found that the introduction of exogenous SCF potently increased KIT kinase activity, stimulated cell proliferation (P < 0.01) and inhibited apoptosis (P < 0.01) induced by serum starvation, while a KIT immunoblocking antibody suppressed proliferation (P = 0.01) and promoted apoptosis (P < 0.01) in cultured GIST cells. CONCLUSION: SCF-mediated wild-type KIT receptor activation plays an important role in GIST cell growth. The inhibition of SCF-mediated wild-type KIT receptor activation may prove to be particularly important for GIST therapy.

pH-Dependence of Manganese (II) Oxidation Reaction by Novel Wild-Type and Mutants Recombinant Phlebia radiata Manganese Peroxidase 3 (rPr-MnP3) Enzymes

The goal of this study was to determine whether mutation of the Mn-binding site of wild-type recombinant Phlebia radiata manganese peroxidase 3 affected the pH-dependence kinetic parameters. pH range investigated was 2.5 – 12.0. The catalytic efficiency of the mutant enzymes at high and low pH in comparison to the wild-type was investigated using standard rPr-MnP3 protocol. Wild-type recombinant Phlebia radiata MnP3 enzyme showed optimal activity with Mn (II) as substrate at pH 5.0 and remained moderately active (approximately 40%) in the pH range of 6.0 - 9.0. The rPr-MnP3 mutants’ maximum activity ranged between 5.5 and 8.0. Wild-type and mutants rPr-MnP3 enzymes exhibited a similar pH profile with optimum pH of 3.0 for ABTS oxidation. Mutation has severely decreased the catalytic efficiency for Mn (II) oxidation at pH 5.0. The rPr-MnP3 enzymes showed enhanced affinity for Mn (II) at alkaline pH and a more alkaline range for catalysis than ever reported for any Manganese Peroxidase. This study reveals that at higher pH, rPr-MnP3 can function with alternative ligands in the Mn (II) site and does not have an absolutely obligate requirement for an all carboxylate ligand set. These results further strongly confirm that Mn2+ binding site is the only productive catalytic site for Mn (II) oxidation.

Case Report: Long-Term Survival in Patient with Cirrhosis of the Liver and Colon Cancer K-ras Wild-Type

K-ras wild-type carcinoma is a tumour that is sensitive to treatment with anti-cancer and anti-EGFR drugs: the combination of Cetuximab and Panitumumab with chemotherapy (Cetuximab) or as a single therapy (Panitumumab). Case Report: The clinical case presented here refers to a 68-year-old patient who had been diagnosed with adenocarcinoma of the recto sigmoid with pelvic recurrence three years after surgery. The patient had a severe co-morbidity: correlated B-type liver cirrhosis. First-line chemotherapy was begun with Oxaliplatin plus Capecitabine (CAPOXI) following a relapse, and this continued for six months (six cycles), when the treatment was interrupted because of the disease’s progression and hematological and gastrointestinal toxicity. Following an assessment of the K-ras, diagnosed as wild type, the patient was excluded from second-line chemotherapy treatment because of decompensated cirrhosis and the persistence of thrombocytopenia and leukopenia. The patient was put forward for biological treatment with an anti-EGFR monoclonal antibody (Panitumumab). Panitumumab was administered at a dosage of 6 mg/kg every 2 weeks for 17 months;the treatment was well tolerated, despite the cirrhosis, and the main toxicity was the skin rash. Conclusion: In patients with severe comorbidities such as cirrhosis of the liver and K-ras wild-type carcinomas, therapy with a monoclonal antibody such as Panitumumab is a treatment that is well tolerated, with few serious toxic side-effects;it also offers advantages in terms of survival and clinical benefits.

Adaptive Response of Wild and Mutant Type Synechococcus cedrorum to a Polychlorinated Pesticide-Endosulfan

The effect of endosulfan, a hexachlorinated pesticide, on growth, inorganic nitrogenous nutrient uptake (NO3-, NO2- and NH4+), change in pigmentation and glycogen content on wild type and chemically mutagenised cells of Synechococcus cedrorum was investigated. The pattern of response to pesticide stress in wild and mutant type was the same. Growth reappeared in both after a period of initial lag in presence of endosulfan. The duration of lag increased with increasing doses of pesticide. Paradoxically, however, the rate of uptake of NO-3, NO-2 and NH+4 pigment and glycogen content progressively increased with increasing doses. The difference in the adaptation response between wild and mutant types was observed only in the concentration of pesticide that could be tolerated; with the mutant tolerating 2.5 fold more.

Therapeutic strategies targeting the epidermal growth factor receptor signaling pathway in metastatic colorectal cancer

More than 1.9 million new colorectal cancer(CRC)cases and 935000 deaths were estimated to occur worldwide in 2020,representing about one in ten cancer cases and deaths.Overall,colorectal ranks third in incidence,but second in mortality.More than half of the patients are in advanced stages at diagnosis.Treatment options are complex because of the heterogeneity of the patient population,including different molecular subtypes.Treatments have included conventional fluorouracil-based chemotherapy,targeted therapy,immunotherapy,etc.In recent years,with the development of genetic testing technology,more and more targeted drugs have been applied to the treatment of CRC,which has further prolonged the survival of metastatic CRC patients.

UE参数、S-TK1、AG490、WIP1诊断TC

目的:探究UE参数、S-TK1、AG490、WIP1对甲状腺癌(TC)的诊断价值。方法:选择我院收治TC患者102例(TC组)和同期收治甲状腺结节良性患者73例(对照组)。比较两组UE参数、S-TK1、AG490、WIP1,分析各指标对TC的诊断价值。结果:TC组的弹性评分、SR值、S-TK1、WIP1水平均高于对照组,AG490低于对照组(P<0.05);ROC曲线分析发现五项指标联合诊断TC的AUC最高(P<0.05)。结论:UE参数与S-TK1、WIP1、AG490联合检测有利于临床诊断TC。

Neuritin野生型蛋白的制备及活性、稳定性的检测及分析

目的制备符合药典结构要求的无标签Neuritin野生型(wild type,WT)蛋白,并完成其存在形式、活性及稳定性鉴定及分析。方法利用基因重组技术构建符合药典结构要求的Neuritin WT酵母重组子,利用SDS-PAGE还原电泳及Western blot对其进行高表达筛选及鉴定;摸索建立无标签Neuritin WT蛋白纯化方法,对纯化后的蛋白进行SDS-PAGE还原电泳分子量鉴定、Western blot免疫原性鉴定、SEC-HPLC精细鉴定及宿主DNA、宿主蛋白残留量检测;利用SDS-PAGE非还原电泳完成存在形式的鉴定;通过参照药典建立的重组Neuritin蛋白活性鉴定方法,检测并分析Neuritin WT纯化蛋白的活性,并观察37℃下不同时间Neuritin WT纯化蛋白的稳定性。结果制备了纯度高达98%以上的Neuritin WT纯化蛋白,蛋白相对分子质量为9.7 kDa,宿主DNA残留量为0.0094 ng/剂,宿主蛋白残留量占比0.0008%;经检测,该纯化蛋白存在单体、二聚体2种形式。活性鉴定结果表明该蛋白具有维持神经元存活的生物学活性;且37℃下3 d内降解率小于10%、7 d内未到达半衰期(蛋白降解率小于35%)。结论成功制备了纯度高达98%、杂质含量及结构符合药典要求的无标签Neuritin WT蛋白,分析了Neuritin WT蛋白的存在形式,并明确了该蛋白具有较好的生物学活性及稳定性,为Neuritin进一步功能、药学研究及生物制品的研制提供了物质基础。

Genetic Evolution Analysis on Wild Isolates of Citrus Tristeza Virus Originated in China Based on Coat Protein Genes Sequences

The coat protein （CP） genes were cloned and sequenced from viral particles of 11 isolates of citrus tristeza virus （CTV） collected from wild citrus plants in China and 4 Chinese isolates from cultivated sweet orange and pummelo varieties, respectively. By analyzing and comparing the nucleotide and amino acid sequences of CP genes, the 11 wild CTV isolates were found over 92% identical with 4 Chinese CTV isolates and 21 exotic CTV isolates from cultivated citrus. From 91 to 100% of the CTV CP gene sequences in wild type citrus plants were generally well conserved. Genetic evolution analysis indicated that the GC% of the CP gene was less than AT%, and more transition were found in the CP genes than transversion with the transition/transversion ratio ranging from 6.3 to 7.0 among species. The substitution frequency was the highest at the third codon, followed by the first and second codon. The ratio of non-synonymous mutations （du） to synonymous mutations （ds） was far lower than 1, suggesting that the CP gene might have experienced purifying selection in the evolution. Phylogenetic analysis revealed that the 11 CTV isolates in Chinese wild type citrus belonged to different phylogenetic clusters, and shared higher homology and closer relationships with other cultivated citrus CTV isolates from different countries, which indicated complicated genetic relationships among the CTV isolates. In addition, CTV isolates with similar biological characteristics usually located into the same clusters. Therefore, the conclusion was drawn that pathogenicity was critical to evolution and origin of CTV.

极小Wild系统箭图代数的Hochschild上同调群

设Γj=kQ/Ij是极小Wild表示型系统箭图代数,基于Bardzell的方法构造了Γj的极小投射双模分解,并由此清晰地计算了Γj的各阶Hochschild上同调群的维数。

沙利度胺联合化疗对EGFR野生型晚期非小细胞肺癌的影响

目的 观察沙利度胺联合化疗治疗表皮生长因子受体(epidermal growth factor receptor, EGFR)野生型晚期非小细胞肺癌(non-small cell lung cancer, NSCLC)患者的生活质量评分、不良反应发生率以及对血清血管内皮生长因子(vascular endothelial growth factor, VEGF)水平的影响。方法 纳入2021年1月至2022年12月在蚌埠医学院第一附属医院肿瘤内科接受治疗的EGFR野生型晚期NSCLC患者,共纳入100例。观察组为采用沙利度胺联合含铂两药化疗治疗患者;对照组为单用含铂两药化疗治疗的患者。观察组共纳入58例患者,给予含铂两药化疗方案治疗;同时口服沙利度胺片100 mg,每日一次,睡前服用;对照组共纳入42例患者,给予化疗方案剂量同上,两组化疗前常规给予止吐药物预防性治疗。比较用药后第7~14天两组患者生活质量各领域评分;比较两组患者治疗期间不良反应发生率;比较两组患者治疗后血清VEGF水平差异。结果 观察组患者第7~14天的躯体(79.18±8.59 vs 64.94±8.60)、角色(71.82±14.05 vs 59.00±12.10)、情绪功能(81.94±15.95 vs 69.71±11.38)、认知(74.53±11.74 vs 59.56±8.75)、社会功能(82.24±13.70 vs 66.82±10.11)和总体健康状况评分(70.81±9.67 vs 60.11±8.77)较对照组患者均明显提高,差异有统计学意义(P<0.05);疲倦(28.08±13.09 vs 38.53±14.23)、疼痛(27.44±8.17 vs 39.19±10.13)、失眠(38.07±12.12 vs 45.23±16.61)和食欲丧失症状评分(40.46±13.70 vs 47.61±17.15)则显著低于对照组,差异有统计学意义(P<0.05);两组间恶心呕吐、气促、便秘、腹泻和经济困难差异无统计学意义(P>0.05);观察组患者化疗期间白细胞及血小板下降不良反应发生率低于对照组(20/58 vs 27/42;P<0.05);两组患者血清VEGF水平比较,观察组水平低于对照组,且差异有统计学意义(127.35±126.03 vs 183.86±120.55;P<0.05)。结论 沙利度胺对EGFR野生型晚期非小细胞肺癌化疗期间的生活质量起到改善作用,且降低不良反应发生率及VEGF水平。

转Pofst3糙皮侧耳菌株农艺性状和胞外酶活性分析

通过袋栽实验比较野生型、Pofst3过表达型和Pofst3反义沉默型糙皮侧耳(Pleurotus ostreatus)菌株(分别记为Ⅰ、Ⅱ、Ⅲ)的菌丝生长速度、原基数量、子实体数量、菌柄直径、菌盖直径等农艺性状;测定其漆酶、木聚糖酶、纤维素酶活性。结果表明:Ⅰ、Ⅱ、Ⅲ菌丝生长速度差异不显著;与Ⅰ相比,Ⅲ的原基和子实体数量多,菌柄直径大,菌盖直径小,原基和子实体中Pofst3的相对表达量低;与Ⅰ、Ⅱ相比,Ⅲ的漆酶、木聚糖酶及纤维素酶活性较高;Pofst3对糙皮侧耳发育的影响可能与其胞外酶活性相关。

EGFR/ALK野生型晚期NSCLC患者血清miR-499 miR-144-3p变化及对预后的预测价值

目的:分析表皮生长因子受体(EGFR)/间变性淋巴瘤激酶(ALK)野生型晚期非小细胞肺癌(NSCLC)患者血清微小RNA(miR)-499、miR-144-3p变化,并评估其对预后的预测价值。方法:收集2018年1月至2020年我院收治的107例EGFR/ALK野生型晚期NSCLC患者临床资料(NSCLC组),另取同期健康体检者107例作为对照组,采用实时荧光RCR技术检测血清miR-499、miR-144-3p表达,比较其在两组中的差异,并评估不同临床病理特征晚期NSCLC患者血清miR-499、miR-144-3p表达的变化;以出院时为随访起点,2023年1月为截止时间,死亡为终点事件,以大于等于平均值作为高表达的标准,分别比较miR-499、miR-144-3p高表达与低表达的晚期NSCLC患者预后生存率。结果:NSCLC组血清miR-499[(0.74±0.18)vs(1.05±0.20)]、miR-144-3p相对表达量[(0.82±0.19)vs(1.22±0.21)]均低于对照组(P<0.05)。晚期NSCLC患者中,Ⅳ期患者血清miR-499[(0.64±0.17)vs(0.86±0.21)]、miR-144-3p相对表达量[(0.71±0.16)vs(0.95±0.22)]低于ⅢB及ⅢC期者(P<0.05),低分化者则低于中高分化者[(0.63±0.16)vs(0.85±0.21),(0.70±0.15)vs(0.94±0.22),P<0.05]。晚期NSCLC患者随访时间为4~56个月,中位随访时间34个月,血清miR-499高表达者生存率明显高于低表达者(P<0.05),血清miR-144-3p高表达者生存率明显高于低表达者(P<0.05)。结论:miR-499、miR-144-3p在EGFR/ALK野生型晚期NSCLC患者血清中呈低表达,Ⅳ期及低分化者表达更低,且表达水平对预后生存率有一定预测价值。

野生越南槐组织特异性内生真菌组及体外抗病原菌功能

有益微生物组能帮助宿主植物防御病害。越南槐的根、茎和种子在野外环境下健康发芽生长,而栽培越南槐各组织极易感病。为了探明利用野生越南槐有益内生真菌组防治宿主病害的可能,该文在分离健康野生越南槐根、茎和种子内生真菌的基础上,并结合形态学和ITS序列特征鉴定内生真菌,通过系统发育树、α-多样性指数和β-多样性指数分别分析各内生真菌组的系统进化、多样性和相似性,通过琼脂块法和平板对峙法测试内生真菌组的体外抗病原菌功能。结果表明:(1)从越南槐根、茎和种子内生真菌组分别分离鉴定131株23个分类单元、108株23个分类单元、64株11个分类单元;(2)特有属多且所有种均为特有种显示根、茎和种子内生真菌组在属种进化上具有组织特异性;(3)根-茎/根-种子/茎-种子间极低的β-多样性显示根、茎和种子各内生真菌组间的物种相似性极低;(4)高的α-多样性显示越南槐根、茎和种子均有丰富多样的内生真菌组;(5)各内生真菌组三分之一以上的分类单元在体外均能拮抗供试病原菌,根茎内生真菌组显示了强广谱的体外抗病原细/真菌功能,种子内生真菌组显示了强广谱的体外抗病原真菌功能。该研究结果表明健康野生越南槐根、茎和种子内部均存在有益内生真菌组,其具有生物多样性、组织特异性以及强广谱的体外抗病原菌功能,在宿主各组织的抗病性方面可能发挥重要作用。这些结果将为进一步利用有益真菌组防治栽培越南槐各组织病害提供了材料和实验基础。