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.

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.

Early and aggressive presentation of wild-type transthyretin amyloid cardiomyopathy:A case report

BACKGROUND Wild-type transthyretin amyloidosis(ATTRwt)is the most common form of transthyretin amyloid cardiomyopathy,occurring mostly over age of 60 years(mean age of 80 years).Mean survival without treatment is 3.6 years,making early detection imperative.We report an unusual case of a 58-year-old patient with ATTRwt cardiomyopathy requiring heart transplantation.CASE SUMMARY A 58-year-old male presented with progressive fatigue,shortness of breath,weight gain,leg swelling,orthopnoea,and paroxysmal nocturnal dyspnoea for several months.Approximately ten months before this clinical presentation,the patient had first received a diagnosis of heart failure with reduced ejection fraction(EF)of 15% to 20%.The patient was started on appropriate guidelinedirected medical therapy with only mild improvement in his EF.Upon further investigation,echocardiogram,technetium pyrophosphate scan(Tc PYP),and cardiac magnetic resonance imaging(cMRI)suggested a diagnosis of amyloidosis,and ATTRwt was subsequently confirmed with native heart tissue biopsy,congo red staining,liquid chromatography-tandem mass spectrometry,and genetic testing.The patient was successfully treated with heart transplantation and is doing well post-transplant.CONCLUSION Wild-type ATTR amyloidosis should be kept on differentials in all patients(even less than 60 years old)with non-ischemic cardiomyopathy,especially in the setting of increased ventricular wall thickness and other classic echocardiogram,cMRI,and Tc PYP findings.Early diagnosis and management can be consequential in improving patient outcomes.

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.

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.

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.

Wild-type p53-induced Phosphatase I Deficiency Exacerbates Myocardial Infarction-induced Ischemic Injury

Background： Myocardial infarction （MI） is a major disease burden. Wild-type p53-induced phosphatase 1 （Wipl） has been studied extensively in the context of cancer and the regulation of different types of stem cells, but the role of Wipl in cardiac adaptation to M I is unknown. We investigated the significance of Wipl in a mouse model of MI. Methods： The study began in June 2014 and was completed in July 2016. We compared Wipl-knockout （Wipl-KO） mice and wild-type （WT） mice to deternline changes in cardiac function and survival in response to MI. The heart weight/body weight （HW/BW） ratio and cardiac function were measured before MI. Mouse MI was established by ligating the left anterior descending （LAD） coronary artery under 1.5% isoflurane anesthesia. After M1, survival of the mice was observed for 4 weeks. Cardiac function was examined by echocardiography. The HW/BW ratio was analyzed, and cardiac hypertrophy was measured by wheat germ agglutinin staining. Hematoxylin and eosin （H＆E） staining was used to determine the infarct size. Gene expression of interleukin-6 （IL-6）, turnor necrosis factor-α （TNF-α）, and interleukin-1β （IL-1β） was assessed by quantitative real-time polymerase chain reaction （qPCR）, and the levels of signal transducers and activators of transcription 3 （stat3） and phosphor-stat3 （p-stat3） were also analyzed by Western blotting. Kaplan-Meier survival analysis, log-rank test, unpaired l-test, and one-way analysis of variance （ANOVA） were used for statistical analyses. Results： Wipl-KO mice had a marginally increased HW/BW ratio and slightly impaired cardiac fiinction before LAD ligation. Alter MI, Wipl-deficient mice exhibited increased mortality （57.14% vs. 29.17%; n = 24 [WT], n - 35 [WipI-KO], P 〈 0.05）, increased cardiac hypertrophy （HW/BW ratio： 7 days： 7.25±0.36 vs. 5.84 ± 0.18, n cross-sectional area： 7 days： 311.80 ± 8.29 vs. 268.90 ± 11.15, n P 〉 0.05）, and reduced cardiac function （ejection fraction： 7 days 10, p〈 0.01, and 4 weeks： 6.05± 0.17 vs. 5.87 ±0.24, n= 10, P〉0.05; P 〈 0.05, and 4 weeks： 308.80 ± 11.26 vs. 317.00 ±13.55, n = 6 29.37± 1.38 vs. 34.72 ± 1.81, P 〈 0.05, and 4 weeks： 19.06 ± 2.07 vs 26.37 ± 2.95, P〈 0.05; fractional shortening： 7 days： 13.72 ± 0.71 vs. 16.50 ± 0.94, P〈 0.05, and 4 weeks： 8.79 ±1.00 vs. 12.48 ±1.48, P 〈 0.05; n = l0 [WT], n = 15 [Wipl-KO]）. H＆E staining revealed a larger infarct size in Wipl-KO mice than in WT mice （34.79% ± 2.44% vs. 19.55% ± 1.48%, n = 6, P 〈 0.01 ）. The expression oflL-6 and p-stat3 was downregulated in Wipl-KO mice （IL-6：1.71 ± 0.27 vs. 4.46 ± 0.79, n = 6, P 〈 0.01 ; and p-stat3/stat3：1.15 ±0.15 vs. 1.97 ± 0.23, n = 6, P 〈 0.05）. Conclusion： The results suggest that Wipl could protect the heart from MI-induced ischemic injury.

A multifunctional nanotheranostic agent potentiates erlotinib to EGFR wild-type non-small cell lung cancer

Epidermal growth factor receptor(EGFR)tyrosine kinase inhibitors(TKI),such as Erlotinib,have demonstrated remarkable efficacy in the treatment of non-small cell lung cancer(NSCLC)patients with mutated EGFR.However,the efficacy of EGFR-TKIs in wild-type(wt)EGFR tumours has been shown to be marginal.Methods that can sensitize Erlotinib to EGFR wild-type NSCLC remain rare.Herein,we developed a multifunctional superparamagnetic nanotheranostic agent as a novel strategy to potentiate Erlotinib to EGFR-wt NSCLCs.Our results demonstrate that the nanoparticles can co-escort Erlotinib and a vascular epithermal growth factor(VEGF)inhibitor,Bevacizumab(Bev),to EGFR-wt tumours.The nanotheranostic agent exhibits remarkable effects as an inhibitor of EGFR-wt tumour growth.Moreover,Bev normalizes the tumour embedded vessels,further promoting the therapeutic efficacy of Erlotinib.In addition,the tumour engagement of the nanoparticles and the vascular normalization could be tracked by magnetic resonance imaging(MRI).Collectively,our study,for the first time,demonstrated that elaborated nanoparticles could be employed as a robust tool to potentiate Erlotinib to EGFR-wt NSCLC,paving the way for imaging-guided nanotheranostics for refractory NSCLCs expressing EGFR wild-type genes.

Emerging role of liquid biopsy in rat sarcoma virus mutated metastatic colorectal cancer:A case report

BACKGROUND In patients with metastatic colorectal cancer(mCRC),the treatment options are limited and have been proved to be affected by rat sarcoma virus(RAS)mutational status.In RAS wild-type(wt)patients,the combination of antiepidermal growth factor receptor(EGFR)monoclonal antibodies with chemotherapy(CT)is more effective than CT alone.On the other hand,RAS-mutated patients are not eligible for treatment with anti-EGFR antibodies.CASE SUMMARY Eleven patients with initially RAS-mutated mCRC were followed from diagnosis to May 2022.At the time of cell-free DNA determination,five patients had undergone one CT line,five patients had undergone two CT lines,and one patient had undergone three CT lines(all in combination with bevacizumab).At the second and third treatment lines[second line(2L),third line(3L)],patients with neo-RAS wt received a combination of CT and cetuximab.In neo-RAS wt patients treated with anti-EGFR,our findings indicated an increase in progression-free survival for both 2L and 3L(14.5 mo,P=0.119 and 3.9 mo,P=0.882,respectively).Regarding 2L overall survival,we registered a slight increase in neo-RAS wt patients treated with anti-EGFR(33.6 mo vs 32.4 mo,P=0.385).At data cut-off,two patients were still alive:A RAS-mutated patient undergoing 3L treatment and a neo-RAS wt patient who received 2L treatment with anti-EGFR(ongoing).CONCLUSION Our case series demonstrated that monitoring RAS mutations in mCRC by liquid biopsy may provide an additional treatment line for neo-RAS wt 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。

卡瑞利珠单抗联合培美曲塞和奈达铂治疗EGFR/ALK野生型晚期非鳞状NSCLC的临床观察

目的观察卡瑞利珠单抗联合培美曲塞和奈达铂治疗表皮生长因子受体(EGFR)和间变性淋巴瘤激酶(ALK)野生型晚期非鳞状非小细胞肺癌(NSCLC)的疗效和安全性。方法回顾性收集2021年8月至2023年5月于重庆医科大学附属第一医院就诊的92例EGFR/ALK野生型晚期非鳞状NSCLC患者资料,根据治疗方案的不同分为奈达铂组(46例)和卡铂组(46例)。奈达铂组患者给予注射用卡瑞利珠单抗+注射用奈达铂+注射用培美曲塞二钠;卡铂组患者给予注射用卡瑞利珠单抗+卡铂注射液+注射用培美曲塞二钠。两组均以21 d为1个周期,所有患者至少完成2个周期的治疗。观察两组患者的近期疗效和不良反应发生情况,分析影响患者无进展生存期(PFS)的因素。结果两组患者的客观缓解率、疾病控制率、中位PFS、3~5级治疗相关不良事件(TRAE)发生率比较,差异均无统计学意义(P>0.05)。奈达铂组患者的1~2级肾脏和泌尿系统TRAE、心慌、心包积液发生率均显著低于卡铂,恶心、呕吐和食欲降低发生率显著高于卡铂组(P<0.05)。患者的性别、年龄、有无吸烟史、美国东部肿瘤协作组织评分和TNM分期均不是影响患者PFS的因素(P>0.05)。结论卡瑞利珠单抗联合培美曲塞和奈达铂治疗EGFR/ALK野生型晚期非鳞状NSCLC的疗效显著,安全性较好。

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。

水疱性口炎病毒对小鼠乳腺癌4T1细胞荷瘤小鼠抗肿瘤免疫、移植瘤生长与肺转移的影响

目的:探究野生型水疱性口炎病毒印第安纳株(VSV-IN)对小鼠三阴性乳腺癌4T1细胞移植模型小鼠的免疫调节及肿瘤转移的影响。方法:VSV以MOI=1、MOI=10、MOI=100感染4T1细胞12、24、48 h后,CCK-8法检测4T1细胞死亡率,划痕愈合实验检测细胞迁移能力,qPCR检测细胞中E-cadherin、MMP-2、MMP-9 mRNA的表达。于雌性BALB/c小鼠脂肪垫接种1×10^(6)个/mL的4T1细胞0.1 mL,构建4T1细胞荷瘤小鼠模型,待小鼠肿瘤体积达200 mm3,分别向移植瘤内注射PBS、紫杉醇(TAX)(15 mg/kg)、VSV-IN(1×10^(6)pfu/只),每周1次。给药4次后,处死小鼠、剥离完整移植瘤组织,测量肿瘤体积及质量,肺组织病理切片经H-E染色后观察肿瘤肺部转移结节,流式细胞术检测脾组织中T细胞亚群比例,ELISA法检测小鼠血清IL-6及TNF-α水平,利用GEPIA在线分析乳腺肿中迁移相关蛋白mRNA的表达,免疫组化法检测肿瘤中MMP-2、MMP-9与E-cadherin的表达,利用蛋白-蛋白对接技术预测VSV-IN的G蛋白、M蛋白与ERK2、E-cadherin的亲和力。结果:经MOI=10、100的VSV-IN处理48 h后,4T1细胞死亡率显著高于对照组(均P<0.01);与对照组相比,VSV-IN组(MOI=10)细胞迁移率明显降低(P<0.01),MMP-9 mRNA的相对表达量明显降低(P<0.05);与对照组小鼠相比,VSV-IN组移植瘤生长较对照组减缓且终点体积显著减小(P<0.05),VSV-IN组小鼠肺转移结节数量显著减少[(12.86±1.86)vs(24±3.67)个,P<0.01],脾内CD4^(+)T、CD8^(+)T细胞比例显著升高(均P<0.05),血清TNF-α、IL-6含量显著升高(均P<0.01);GEPIA分析发现在乳腺癌中E-cadherin、MMP-9表达水平均高于癌旁组织(P<0.05);VSV-IN组小鼠肿瘤细胞内MMP-9表达明显低于对照组(P<0.05);VSV-IN的G、M蛋白与ERK2的结合自由能分别为–11.7 kcal/mol、–6.4 kcal/mol。结论:野生型VSV-IN可抑制4T1细胞荷瘤小鼠的移植瘤生长及转移,这可能与其促进抗肿瘤免疫及调控迁移相关蛋白表达有关。

基于机器学习方法构建IDH野生型胶质母细胞瘤预测模型研究

目的建立异柠檬酸脱氢酶(IDH)野生型胶质母细胞瘤生存概率的列线图模型及随机生存森林模型。方法回顾性分析2017年1月—2020年12月在空军军医大学附属西京医院手术治疗的127例IDH野生型胶质母细胞瘤患者临床资料,进行预后因素分析并建立列线图模型及随机生存森林模型,通过C指数,校准曲线,决策曲线评价模型的区分度,校准度以及临床净获益率。结果使用Cox比例风险模型进行多因素分析发现,患者术前卡氏功能状态评分(KPS)、是否接受同步放化疗、年龄、O^(6)-甲基鸟嘌呤甲基转移酶(MGMT)蛋白表达,是独立的预后因素(P<0.05)。通过Cox回归模型建立列线图预测模型;通过R软件建立随机生存森林模型,两个模型均具有良好的区分度和校准度,随机生存森林模型的临床净获益优于列线图模型。结论建立的列线图模型及随机生存森林模型有助于临床医生判断患者特定时间点的生存概率。

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。