Pancreatic neuroendocrine tumor detected by technetium-99m methoxy-2-isobutylisonitrile single photon emission computed tomography/computed tomography:A case report

BACKGROUND Pancreatic neuroendocrine tumors(NETs)account for about 1%–2%of pancreatic tumors and about 8%of all NETs.Computed tomography(CT),magnetic resonance imaging,and endoscopic ultrasound are common imaging modalities for the diagnosis of pancreatic NETs.Furthermore,somatostatin receptor imaging is of great value for diagnosing pancreatic NETs.Herein,we report the efficacy of technetium-99m methoxy-2-isobutylisonitrile(99mTc-MIBI)single photon emission CT(SPECT)/CT for detecting pancreatic NETs.CASE SUMMARY A 57-year-old woman presented to our hospital with a 1-d history of persistent upper abdominal distending pain.The distending pain in the upper abdomen was aggravated after eating,with nausea and retching.Routine blood test results showed a high neutrophil percentage,low leukomonocyte and monocyte percentages,and low leukomonocyte and eosinophil counts.Amylase,liver and kidney function,and tumor markers alpha-fetoprotein,carcinoembryonic antigen,and cancer antigen(CA)125,CA72-4,CA19-9,and CA153 were normal.Abdominal CT showed a mass,with multiple calcifications between the pancreas and the spleen.The boundary between the mass and the pancreas and spleen was poorly defined.Contrast-enhanced CT revealed that the upper abdominal mass was unevenly and gradually enhanced.99mTc-MIBI SPECT/CT revealed that a focal radioactive concentration,with mild radioactive concentration extending into the upper abdominal mass,was present at the pancreatic body and tail.The 99mTc-MIBI SPECT/CT manifestations were consistent with the final pathological diagnosis of pancreatic NET.CONCLUSION 99mTc-MIBI SPECT/CT appears to be a valuable tool for detecting pancreatic NETs.

Direct technetium-99m labeling of anti-hepatoma monoclonal antibody fragment:a radioimmunoconjugate for hepatocellular carcinoma imaging

AIM To directly radiolabel an anti-hepatomamAb fragment HAb18 F（ab’）2 with 99mTc bystannous-reduced method,and assess thestability,biodistribution and radioimmun-oimaging（RⅡ）.METHODS Immunoreactive fraction wasdetermined according to Lindmo’s method.Ellman’s reagent was used to determine thenumber of thiols in the reduced F（ab’）2.Labelingefficiency and homogeneity were measured bypaper chromatography,sodium dodecylsulphatepolyacrylamide gel electrophoresis（SDS-PAGE）and autoradiography.Challenge assay involvedthe incubation of aliquots of labeled antibody inethylenediaminetetraacetate（ EDTA ）and L-cysteine（L-cys）solutions with different molarratio at 37℃ for 1h,respectively.Investigationsin vivo utilized nude mice bearing humanhepatocellular carcinoma（HHCC）xenograftswith gamma camera imaging and tissuebiodistribution studies at regular intervals.RESULTS The labeling procedure was finishedwithin 1.5 h compared with the'pretinning'method which would take at least 21h.In vitrostudies demonstrated that the radiolabeled mAbfragment was homogeneous and retained itsimmunoreactivity.Challenge studies indicatedthat 99mTc-labeled HAb18 F（ab’）2 in EDTA is morestable than in L-cys.Imaging and biodistribution showed a significant tumor uptake at 24 h post-injection of 99mTc-labeled HAb18 F（ab’）2.Theblood,kidney,liver and tumor uptakes at 24hwere 0.56±0.09,56.45±11.36,1.43±0.27 and6.57±3.01（%ID/g）,respectively.CONCLUSION 99mTc-HAb18 F（ab’）2 conjugateprepared by this direct method appears to be aneffective way to detect hepatoma in nude micemodel.

Effect of Technetium-99 Conjugated with Methylene Diphosphonate on IgM-RF,IgG-RF and IgA-RF

To explore the effect of technetium-99 conjugated with methylene diphosphonate ( 99 Tc-MDP) on IgM-RF, IgG-RF and IgA-RF (RFs), 47 cases were selected for study, including 33 patients with rheumatoid arthritis (RA) and 15 patients with joint pain/arthritis. After 99 Tc-MDP for drips model being given to the patients by intravenous drip 0.2 g daily for 5 days, the injection A and B models of 99 Tc-MDP were used to the patients by intravenous injection one set daily for 10 days, that was one course of treatment. The next course started after 10 days. Each case used it from 2 to 4 courses of treatment. The RFs in serum were determined by the method of enzyme-linked immunoabsorption assay (ELISA) before and after 2 and 4 courses of treatment. In the patients with RA, the concentrations of IgM-RF were 296.2±108.4 IU/ml, 189.5±92.3 IU/ml and 107.8±72.5 IU/ml; the concentrations of IgG-RF were 325.6±126.2 IU/ml, 209.7±98.2 IU/ml and 160.2±80.8 IU/ml; the concentrations of IgA-RF were 330.4±136.3 IU/ml, 210.7±89.2 IU/ml and 148.8±72.2 IU/ml before and after 2 and 4 courses of treatment, respectively. The concentrations of the above RFs were significantly lower after 2 and 4 courses than those before treatment ( P <0.05 and P <0.01). There was no significant difference in RFs concentrations in the patients with joint pain/arthritis before and after use of 99 Tc-MDP. In the patients with positive RFs before treatment, the RFs concentrations were decreased significantly after 2 and 4 courses of treatment ( P <0.05 and P <0.01). There was no obvious change of RFs concentrations in the patients with negative RFs after treatment of 99 Tc-MDP. It was concluded that 99 Tc-MDP could obviously reduce the abnormally high concentrations of RFs, but not influence the normal RFs, which indicated that 99 Tc-MDP has an important effect on controlling the activities of RA.

Technetium-99m-labeled macroaggregated albumin lung perfusion scan for diagnosis of hepatopulmonary syndrome: A prospective study comparing brain uptake and whole-body uptake

BACKGROUND Hepatopulmonary syndrome (HPS) is an arterial oxygenation defect induced by intrapulmonary vascular dilatation (IPVD) in the setting of liver disease and/or portal hypertension.This syndrome occurs most often in cirrhotic patients(4%-32%) and has been shown to be detrimental to functional status,quality of life,and survival.The diagnosis of HPS in the setting of liver disease and/or portal hypertension requires the demonstration of IPVD (i.e.,diffuse or localized abnormally dilated pulmonary capillaries and pulmonary and pleural arteriovenous communications) and arterial oxygenation defects,preferably by contrast-enhanced echocardiography and measurement of the alveolar-arterial oxygen gradient,respectively.AIM To compare brain and whole-body uptake of technetium for diagnosing HPS.METHODS Sixty-nine patients with chronic liver disease and/or portal hypertension were prospectively included.Brain uptake and whole-body uptake were calculated using the geometric mean of technetium counts in the brain and lungs and in the entire body and lungs,respectively.RESULTS Thirty-two (46%) patients had IPVD as detected by contrast-enhancedechocardiography.The demographics and clinical characteristics of the patients with and without IPVD were not significantly different with the exception of the creatinine level (0.71±0.18 mg/dL vs 0.83±0.23 mg/dL;P=0.041),alveolararterial oxygen gradient (23.2±13.3 mmHg vs 16.4±14.1 mmHg;P=0.043),and arterial partial pressure of oxygen (81.0±12.1 mmHg vs 90.1±12.8 mmHg;P=0.004).Whole-body uptake was significantly higher in patients with IPVD than in patients without IPVD (48.0%±6.1%vs 40.1%±8.1%;P=0.001).The area under the curve of whole-body uptake for detecting IPVD was significantly higher than that of brain uptake (0.75 vs 0.54;P=0.025).The optimal cut-off values of brain uptake and whole-body uptake for detecting IPVD were 5.7%and 42.5%,respectively,based on Youden’s index.The sensitivity,specificity,and accuracy of brain uptake> 5.7%and whole-body uptake> 42.5%for detecting IPVD were23%,89%,and 59%and 100%,52%,and 74%,respectively.CONCLUSION Whole-body uptake is superior to brain uptake for diagnosing HPS.

Leukocyte-technetium-99m uptake in Crohn's disease:Does it show subclinical disease?

AIM:To evaluate inflammatory activity in patients with Crohn's disease (CD) using technetium-99m-hexamethylpropyleneamine oxime (99mTc-HMPAO) granulocyte scintigraphy.METHODS: Twenty patients (7 male and 13 female) with CD and five healthy volunteers were selected for 99mTc-HMPAO granulocyte scintigraphy. The Crohn's Disease Activity Index (CDAI), blood tests and C-reactive protein (CRP) of each patient were performed 7 d before the scintigraphic images. The leukocytes were labeled according to the International Society of Radiolabeled Blood Elements (ISORBE) consensus protocol and the scintigraphic images, including single photon emission computed tomography, were obtained 30 min and 2 h after injection of the radiolabeled leukocytes.RESULTS:The labeling yield of the leukocytes with the lipophilic complex 99mTc-HMPAO was 55.0%±10%. Six of the 20 patients (30%) presented congruent results for the three parameters investigated (CDAI, Scintigraphic Index and CRP). On the other hand, 14 patients (70%) did not show congruent results. There was no significant correlation between the indices analyzed according to the Spearman test (P>0.05,n=20).CONCLUSION: The results suggest that 99mTc-HMPAO-labeled leukocyte scintigraphy could be important for determining inflammatory activity in CD even in the absence of clinical symptoms.

Technetium-99m-labeled annexin V imaging for detecting prosthetic joint infection in a rabbit model

Accurate and timely diagnosis of prosthetic joint infection is essential to initiate early treatment and achieve a favorable outcome. In this study, we used a rabbit model to assess the feasibility of technetium-99m-labeled annexin V for detecting prosthetic joint infection. Right knee arthroplasty was performed on 24 New Zealand rabbits. After surgery, methicillin-susceptible Staphylococcus aureus was intra-articularly injected to create a model of prosthetic joint infection （the infected group, n = 12）. Rabbits in the control group were injected with sterile saline （n= 12）. Seven and 21 days after surgery, technetium-99m-labeled annexin V imaging was per- formed in 6 rabbits of each group. Images were acquired 1 and 4 hours after injection of technetium-99m- labeled annexin V （150 MBq）. The operated-to-normal-knee activity ratios were calculated for quantitative ana- lysis. Seven days after surgery, increased technetium-99m-labeled annexin V uptake was observed in all cases. However, at 21 days a notable decrease was found in the control group, but not in the infected group. The operated-to-normal-knee activity ratios of the infected group were 1.84 ±0.29 in the early phase and 2.19 ±0.34 in the delay phase, both of which were significantly higher than those of the control group （P=0.03 and P=0.02）. The receiver operator characteristic curve analysis showed that the operated-to-normal-knee activity ratios of the delay phase at 21 days was the best indicator, with an accuracy of 80%. In conclusion, technetium- 99m-labeled annexin V imaging could effectively distinguish an infected prosthetic joint from an uninfected prosthetic joint in a rabbit model.

A NEW BIFUNCTIONAL CHELATING AGENT α,ε-N,N'-BIS(L-CYSTEINYL)-L-LYSINE FOR RADIOLABELLING OF MONOCLONAL ANTIBODIES WITH TECHNETIUM-99M

α,ε-N,N'-bis(L-cysteinyl)-L-lysine was synthesized and char- acterized for the first time.It was then employed as a bifunctional chelating agent to chelate technetium-99m and subsequently conjugated to fragment F(ab')_2 of anti-gastric tumor monoclonal antibody 3G9.The radiolabelled antibody was satisfactorily stable and immunoreactive.

Anti-sense oligonucleotide labeled with technetium-99m using hydrazinonictinamide derivative and N-hydroxysuccinimidyl S-acetylmercaptoacetyltriglycline: A comparison of radiochemical behaviors and biological properties

AIM:To explore and compare the radiochemical behavior and biological property of anti-sense oligonuc-leotide (ASON) labeled with technetium-99m using N-hydroxysuccinimidyl S-acetylmercaptoacetyltriglycl ine (NHS-MAG3) and hydrazinonictinamide derivative (HYNIC). METHODS:After HYNIC and NHS-MAG3 were synthesized, ASON was labeled with technetium-99m using HYNIC and NHS-MAG3 as a bifunctional chelator. The in vivo and in vitro stability, binding rates of labeled compounds to serum albumen, biodistribution of 99mTc-MAG3-ASON and 99mTc-HYNIC-ASON in BALB/C mouse and its HT29 tumor cellular uptake were compared. RESULTS:The labeling efficiency and stability of 99mTc-MAG3-ASON were significantly higher than those of 99mTc-HYNIC-ASON (P = 0.02, and P = 0.03, respectively). 99mTc-MAG3-ASON had a significantly lower rate of binding to serum albumen than 99mTc-HYNIC-ASON (P < 0.05). In contrast to 99mTc-HYNIC-ASON, the biodistribution of 99mTc-MAG3-ASON was significantly lower in blood, heart, liver and stomach (P < 0.05), slightly lower in intestines and spleen (P > 0.05) and significantly higher in lung and kidney (P < 0.05). The HT29 tumor cellular uptake rate of 99mTc-MAG3-ASON was significantly higher than that of 99mTc-HYNIC-ASON (P < 0.05). CONCLUSION:99mTc-MAG3-ASON shows superior radiochemical behaviors and biological properties than 99mTc-HYNIC-ASON. 99mTc-MAG3-ASON is a potential radiopharmaceutical agent for in vivo application.

Incidental accumulation of Technetium-99m pertechnetate in subacute cerebral infarction: A case report

BACKGROUND When interpreting nuclear medicine images,unexpected findings are sometimes encountered.Recognizing these findings and determining the mechanism of their occurrence could have a significant impact on early diagnosis of critical diseases and the appropriate management of patients.CASE SUMMARY A 59-year-old man was admitted to the emergency room due to left hemiparesis,left hemifacial palsy,and mild dysarthria.After 2 wk of hospitalization,the patient complained of dry eyes and mouth.Thus,salivary scintigraphy was performed to evaluate the functional status of his salivary glands.Incidental accumulation in the right frontoparietal area was found on salivary scintigraphy.Fluid-attenuated inversion recovery phase magnetic resonance(FLAIR phase MR)image showed diffuse high signal intensity in the same area.Anterior and posterior horns of the right lateral ventricle were obliterated and the midline was slightly shifted to the left side due to right frontoparietal swelling.On salivary scintigraphy,Tc-99m pertechnetate was incidentally accumulated in a subacute cerebral infarction lesion.Two years after the diagnosis of acute infarction,the second series of salivary scintigraphy showed no abnormal activity in the brain.FLAIR phase MR image also demonstrated markedly decreased high signal intensity in the previous infarction lesion without evidence of swelling indicating chronic cerebral infarction.CONCLUSION This case highlights that Tc-99m pertechnetate could accumulate in a subacute cerebral infarction lesion.The mechanism of an unexpected uptake of Tc-99m pertechnetate in unusual sites should be evaluated and kept in mind for better interpretation.

Preliminary clinical application of dopamine transporter imaging with technetium-99m TRODAT-1 and SPECT on the early and differential diagnosis of Parkinson's Disease

The aim of the study was to demonstrate the degeneration of the dopaninergic nigrostriatal pathway in Parkinson's disease(PD) and Essential Tremor (ET) by using the cocaine derivative 99mTc-TRODAT-1 SPECT and correlate the findings to the clinical severities (Hoehn and Yahr scale, H/Y). 28 patients with idiopathic Parkinson's disease, 10 patients with Essential Trenor and 19 healthy volunteers were investigated. The acquisition were performed 3 h postinjection of 99mTc-TRODAT-1, ROIs were drawn over the images of striatum and cerebellum, and ratios of striatum to cerebellar(ST/CB) were calculated. Ratios differed significantly between PD and controls, but ratios didn't show significant difference between ET patients and controls. A significant correlation didn't exist between ratios and clinical severities. Hemiparkinson's patients revealed significantly diminished 99mTc-TRODAT-1 binding not only clinically affected but unaffected side. Our findings indicated that 99mTc-TRODAT-1 SPECT is not only a reliable method to discriminate between PD and controls but also a useful tool for differential diagnosis in clinically unclear cases such as ET resembling PD.

医用同位素^(99)Mo吸附分离研究进展

医用同位素^(99)Mo是一种广泛应用于核医学领域的重要核素。由于常规的高浓缩铀裂变生产^(99)Mo的过程中存在安全隐患,人们已经开始寻找其他可靠的^(99)Mo生产途径。在分离^(99)Mo和^(99)mTc的方法中柱层析法具有很大优势,其中的关键是层析柱的材料,材料对^(99)Mo吸附能力关系到未来新一代^(99)Mo-^(99)mTc发生器的制备。本研究对医用同位素^(99)Mo的吸附分离进行综述,介绍^(99)Mo生产方式,^(99)Mo和^(99)mTc分离方法,以及目前对Mo具有一定吸附效果的吸附材料,为未来利用低比活度^(99)Mo吸附制备^(99)Mo-^(99)mTc发生器提供参考。

^(99m)Tc-MIBI门控心肌灌注显像对行沙库巴曲缬沙坦治疗的冠心病患者心肌血流灌注及心功能的评估价值

目的:探究^(99m)Tc-MIBI门控心肌灌注显像(GMPI)评估冠心病患者沙库巴曲缬沙坦治疗后心肌血流灌注及心功能的价值。方法:选取冠心病患者78例,入组患者均接受沙库巴曲缬沙坦治疗,采用^(99m)Tc-MIBI GMPI对患者的心肌血流灌注情况及心功能指标进行检测,并以超声心动图及冠状动脉造影结果为参照,对^(99m)Tc-MIBI GMPI检查的准确性进行探究。结果:患者经冠状动脉造影检查结果与患者经^(99m)Tc-MIBI GMPI检查心肌血流灌注结果比较无统计学差异(P>0.05),经卡帕一致性检验,KAPPA值为0.826,一致性较好。患者经冠状动脉造影检查结果与患者经^(99m)Tc-MIBI GMPI心功能结果[舒张末期容积(EDV)、收缩末期容积(ESV)、左心室射血分数(LVEF)、心输出量(CO)]比较无统计学差异(均P>0.05),经ICC一致性检验,ICC=0.812,一致性较好。治疗后与治疗前比较,患者的心肌血流灌注有所好转,患者完全闭塞、重度狭窄占比降低,轻度狭窄占比升高(均P<0.05)。与治疗前比较,治疗后患者的EDV、ESV水平降低,LVEF、CO升高(均P<0.05)。78例患者经过治疗,出现8例症状性低血压,2例轻度血管性水肿,6例肾功能损伤,3例高钾血症,并发症发生率约为24.36%。结论:^(99m)Tc-MIBI GMPI评估心肌血流灌注及心功能情况与超声心动图及冠状动脉造影结果相一致,可以准确地反映冠心病患者经沙库巴曲缬沙坦治疗后的心肌血流灌注及心功能变化。经沙库巴曲缬沙坦治疗,患者的心肌血流灌注及心功能得到改善,但需要对患者的不良反应情况进行关注。

^(99m)TcO4-SPECT/CT甲状腺核素显像联合超声及TSH对甲状腺结节性质的诊断价值

目的通过高锝酸钠-单光子发射计算机断层显像/电子计算机X射线断层扫描技术(^(99m)TcO4-SPECT/CT)核素显像提供甲状腺结节功能状态、结节解剖特点以及超声检查联合血清促甲状腺激素(TSH)水平进一步鉴别诊断甲状腺结节的良恶性,客观评价甲状腺结节的良恶性情况,对甲状腺结节的良恶性做出早期诊断。方法对因甲状腺结节同时行^(99m)TcO4-SPECT/CT甲状腺核素显像、甲状腺B超及TSH水平检查的患者共80例(92个结节)进行回顾性分析,对比不同方法及联合检查对甲状腺结节的诊断效能。结果92个结节中良性74个,恶性18个;^(99m)TcO4-SPECT/CT甲状腺显像与超声检查比较灵敏度(77.8%)高,特异度(70.2%)及准确度(71.7%)低,假阳性率(29.7%)高,假阴性率(22.2%)低,两者之间灵敏度、特异度、准确度、假阳性率、假阴性率差异均具有统计学意义(P<0.05);两者准确度比较差异无统计学意义(P>0.05)。^(99)mTcO4-SPECT/CT甲状腺显像与超声检查比较灵敏度高,特异性及准确度低,假阳性率高,假阴性率低,两者之间灵敏度、特异度、准确度、假阳性率、假阴性率差异均具有统计学意义(P<0.05);两者准确度比较差异无统计学意义(P>0.05);^(99m)TcO4-SPECT/CT甲状腺显像与超声检查联合较单独核素、单独超声检查灵敏度(88.9%)、特异度(87.8%)、准确度(81.5%)高,假阳性率(12.1%)、假阴性率(11.1%)低;联合检查与单独核素特异度、准确度、假阳性率比较差异有统计学意义(P<0.05),灵敏度与假阴性率比较差异无统计学意义(P>0.05);联合检查与单独超声灵敏度、准确度、假阴性率比较差异有统计学意义(P<0.05);特异度与假阳性率比较差异无统计学意义(P>0.05)。结论超声可以作为甲状腺结节的首选检查方法,其特异性高,但灵敏度低。^(99m)TcO4-SPECT/CT甲状腺显像在提供功能和代谢信息的同时,能提供解剖信息;而血清TSH水平则可为甲状腺结节的良恶性鉴别提供重要的参考依据,尤其对于核素冷结节且超声实性结节的患者,更应参考其TSH水平,将三者结合起来判断甲状腺结节的性质,为临床提供恰当的治疗方案,这具有重要意义。

RNF99通过TAK1/NF-κB信号通路参与泛素化与脓毒症性休克的潜在联系

目的 探讨环指蛋白99(RNF99)介导的转化生长因子激酶1(TAK1)/核因子-κB(NF-κB)信号通路参与泛素化与脓毒症性急性呼吸窘迫综合征(ARDS)的潜在联系。方法 进行质粒和siRNA转染以过表达或敲低小鼠肺泡上皮细胞(MLE12)中RNF99,分析磷酸p65和p65蛋白表达。免疫沉淀分析RNF99与TRAF6和TAK1的蛋白相互作用关系。将40只小鼠随机分成WT+PBS、WT+LPS、RNF99特异性表达(TG)+PBS和TG+LPS组,每组10只。通过腹膜内注射30 mg/kg LPS诱导脓毒症。结果 与Vector组相比,RNF99组MLE12细胞中TRAF6和TAK1的蛋白表达水平显著降低(P<0.05)。泛素化TRAF6蛋白在RNF99敲低的MLE12细胞中增加。与LPS+Vector组相比,在LPS+RNF99组MLE12细胞中p65的磷酸化水平明显降低(P <0.05)。与si-NC组相比,si-RNF99组MLE12细胞中RNF99、IκBα的蛋白表达水平显著降低(P <0.05)。与LPS+si-NC组相比,在LPS+si-RNF99组MLE12细胞中p65的磷酸化水平明显增加(P <0.05)。TG+LPS组小鼠肺组织中CD68巨噬细胞染色百分比较WT+LPS组显著降低(P <0.05)。TG+LPS组小鼠肺组织中p65的磷酸化水平显著低于WT+LPS组小鼠(P <0.05)。结论 RNF99通过与NF-κB信号通路的关键调节因子(TRAF6/TAK1)相互作用来调节NF-κB信号通路,并改善小鼠腹腔注射LPS后肺损伤。

靶向CD99的CAR-T细胞扩增优化研究

背景与目的:嵌合抗原受体T(chimeric antigen receptor T,CAR-T)细胞疗法对血液和淋巴系统肿瘤的治疗效果显著,但对实体瘤效果较差,这与靶点选择的因素有关。针对尤因肉瘤(Ewing sarcoma,ES),CD99可作为CAR-T细胞潜在的靶点。由于T细胞自身表达CD99蛋白,靶向CD99的CAR-T细胞存在体外扩增能力有限的问题。本研究旨在通过增加CD99敲低的短发夹RNA(short hairpin RNA,shRNA)、优化慢病毒转导的感染复数(multiplicity of infection,MOI)、筛选培养CAR-T细胞的培养基及培养容器,以获得CD99 CAR-T细胞制备的最优条件。方法:筛选shRNA序列,提高CD99 CAR-T细胞的扩增能力。采用不同的MOI、培养基和培养容器,分别检测在不同条件下CAR-T细胞的转导效率、细胞存活率、增殖能力、特异性杀伤能力及干扰素-γ(interferon-γ,IFN-γ)释放水平等,筛选出最优的细胞制备条件。结果:通过shRNA敲低得到的KO-CD99 CAR-T细胞扩增水平显著高于CD99 CAR-T细胞[(16.40±0.40)vs(6.33±1.53),P<0.01]。转导MOI为0.25~1.0、培养基为OptiVitro时细胞的扩增效果最优。在透气瓶中培养的CAR-T细胞扩增倍数显著高于在培养袋中培养的细胞[MOI=0.25:(50.23±3.32)vs(13.02±4.82);MOI=0.50:(49.96±0.83)vs(18.25±2.88);MOI=1.00:(48.27±5.08)vs(13.16±6.26);P<0.01],且细胞分型更优、特异性杀伤更高。结论:通过shRNA技术得到的KO-CD99 CAR-T细胞可实现稳定扩增。从扩增条件优化结果看,MOI为0.25~1.00,培养基为OptiVitro,培养容器为透气瓶的条件下,KO-CD99 CAR-T细胞可获得更优的扩增能力、更多比例的记忆T细胞,为后续开展CD99 CAR-T细胞治疗ES的临床试验奠定了坚实基础。

猪ETECK88ac、K99黏附素蛋白生物信息学分析及多表位疫苗载体构建

由猪产肠毒素大肠杆菌(ETEC)引起的仔猪腹泻是规模化猪场最常见的疾病之一。目前国内和国际上制备ETEC疫苗的靶基因多为其黏附素基因K88ac、K99。本试验通过分析ETEC K88ac、K99黏附素蛋白的生物学信息,将抗原表位进行组合,设计了一种同时含有K88ac、K99基因的多表位重组疫苗载体。首先应用ProParam、SOPMA和GOR软件分析K88ac、K99蛋白的理化特性及二级结构,应用IEDB与ABCpred软件预测二者的B细胞抗原表位,应用RANKPEP软件预测Th细胞表位,应用IEDB与NetMHC-4.0软件预测CTL细胞表位;然后将预测的B细胞、Th细胞和CTL细胞抗原表位加上接头序列,按照一定顺序进行组合,分别设计多表位连接多肽Ⅰ、Ⅱ和Ⅲ;最后通过分析抗原性、致敏性,筛选抗原性最强的连接多肽,并对其进行表征。结果显示:共筛选出9个抗原表位,按照不同的排列组合获得了3种连接多肽,其中连接多肽Ⅱ抗原性最强,为0.906 8;应用AllerTOP v. 2.0在线工具对连接多肽Ⅱ进行过敏性预测,发现其没有过敏性;应用ZpGJn4在线工具对连接多肽Ⅱ进行三维建模,发现其结构表位暴露性较好,易与抗体结合,在蛋白分子构象上符合表位设计要求。将该多表位肽的氨基酸序列按照乳酸菌密码子嗜好性反向翻译为核苷酸序列,随后插入pET28a载体中获得了p ET28a-KT质粒。结果表明,构建的连接多肽Ⅱ适合作为多表位疫苗的备选载体蛋白。多表位肽的设计及pET28a-KT质粒的获得为下一步构建表达ETEC多表位抗原的重组乳酸菌提供了技术支撑。

锝[^(99)Tc]亚甲基二膦酸盐注射液联合甲氨蝶呤及硫酸羟氯喹治疗对类风湿性关节炎患者红细胞沉降率及类风湿因子的影响

目的:探究锝[^(99)Tc]亚甲基二膦酸盐注射液联合甲氨蝶呤及硫酸羟氯喹治疗对类风湿性关节炎(RA)患者红细胞沉降率(ESR)及类风湿因子(RF)的影响。方法:选取2021年4月—2022年5月抚州市立医院收治的RA患者66例,选用随机数字表法分为两组,各33例。对照组接受甲氨蝶呤及硫酸羟氯喹治疗,观察组接受锝[^(99)Tc]亚甲基二膦酸盐注射液联合甲氨蝶呤及硫酸羟氯喹治疗,两组均持续治疗3个月。对比两组临床疗效、炎症因子水平、ESR及RF、类风湿关节炎活动度(DAS28)评分、症状改善情况、视觉模拟评分法(VAS)评分、不良反应。结果:观察组临床总有效率为96.97%(32/33),高于对照组的75.76%(25/33),差异有统计学意义(P<0.05);治疗后,观察组白介素-6(IL-6)(3.52±0.47)pg/mL,肿瘤坏死因子-α(TNF-α)(3.91±1.02)pg/mL,C蛋白反应(CRP)(12.33±5.15)mg/L,RF(31.62±15.16)IU/mL,ESR(25.89±10.69)mm/h,均低于对照组的(8.03±1.02)pg/mL、(7.94±2.54)pg/mL、(16.38±6.41)mg/L、(42.98±16.87)IU/mL、(35.92±11.84)mm/h,关节肿胀数目(2.01±0.45)个,关节压痛数目(4.15±1.31)个,均少于对照组的(4.05±1.56)、(6.14±2.02)个,僵硬时间(25.56±9.13)min,短于对照组的(32.85±11.85)min,双手平均握力(12.03±2.91)kPa,高于对照组的(9.51±1.05)kPa,DAS28评分(2.02±0.74)分,VAS评分(2.12±0.29)分,均低于对照组的(4.38±1.29)、(4.15±0.57)分,差异均有统计学意义(P<0.05);两组不良反应发生率比较,差异无统计学意义(P>0.05)。结论:锝[^(99)Tc]亚甲基二膦酸盐注射液联合甲氨蝶呤及硫酸羟氯喹在RA患者中应用效果较佳,能够减轻炎症反应,改善临床症状及关节活动度,减轻疼痛,且未增加不良反应。

疑诊原发性甲状旁腺功能亢进症的患者术前^(99m)Tc-MIBI SPECT/CT显像结果的影响因素分析

目的探讨疑诊原发性甲状旁腺功能亢进症(primary hyperparathyroidism,PHPT)的患者术前^(99m)Tc-甲氧基异丁基异腈(methoxyisobutylisonitrile,MIBI)单光子发射计算机断层扫描/计算机断层扫描(single-photon emission computed tomography/computed tomography,SPECT/CT)(以下简称MIBI SPECT/CT)显像结果的影响因素。方法将2019年1月至2023年8月于首都医科大学附属北京朝阳医院疑诊PHPT并在术前完善甲状旁腺MIBI SPECT/CT显像的257例患者作为研究对象。比较MIBI SPECT/CT显像阳性组和阴性组的实验室检查、病理特征及影像特征。采用多因素Logistic回归分析MIBI SPECT/CT显像结果的影响因素。结果单因素分析显示,与阴性组相比,血清总钙、血清甲状旁腺激素(parathyroid hormone,PTH)、碱性磷酸酶(alkaline phosphatase,ALP)水平在阳性组显著升高(P<0.05);病灶的病理类型、最大径、体积及位置在两组间差异有统计学意义(P<0.05)。多因素Logistic回归分析显示,术前血清PTH水平、病灶病理类型、位置及最大径与MIBI SPECT/CT显像结果阳性显著相关(P<0.05)。结论疑诊PHPT患者术前MIBI SPECT/CT显像结果受多重因素影响,术前血清PTH水平、病灶病理类型、位置及最大径是MIBI SPECT/CT显像结果的独立影响因素。

^(99)锝-亚甲基二膦酸盐注射液、唑来膦酸和鲑降钙素在绝经后骨质疏松症患者治疗中的应用比较

目的对比^(99)锝-亚甲基二膦酸盐注射液(^(99)Tc-MDP)、唑来膦酸注射液和鲑降钙素注射液治疗绝经后骨质疏松症(OP)患者的临床疗效及对骨转换指标、炎症因子、骨密度的影响。方法将2021年3月至2023年3月厦门医学院附属第二医院收治的75例绝经后OP患者纳入本次前瞻性研究,并采用随机数字表法将患者分为观察1组、观察2组和观察3组,每组各25例。3组患者均给予骨化三醇和碳酸钙D3治疗,此外,观察1组患者给予^(99)Tc-MDP治疗,观察2组患者给予唑来膦酸注射液治疗,观察3组患者给予鲑降钙素注射液治疗。3组疗程均为3个月。比较3组治疗3个月后的血清I型原胶原N-端前肽(TP1NP)、Ⅰ型胶原C-末端肽交联(β-CTX)、血清炎症因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)]水平,治疗1、3和6个月后疼痛数字分级评分法(NRS)评分,治疗12个月后的腰椎、全髋和股骨颈骨密度及不良反应发生情况。结果3组患者治疗3个月后的TP1NP、β-CTX、TNF-α、IL-6水平比较,差异均有统计学意义(P<0.05),观察1组和观察3组治疗3个月后的TP1NP、β-CTX水平均显著高于观察2组,而TNF-α、IL-6水平均显著低于观察2组,差异均有统计学意义(P<0.05),观察1组和观察3组治疗3个月后的TP1NP、β-CTX、TNF-α、IL-6水平比较,差异均无统计学意义(P>0.05)。3组治疗1、3和6个月的NRS评分比较,差异均有统计学意义(P<0.05),观察1组和观察3组治疗1、3和6个月后的NRS评分均显著低于观察2组,而观察1组和观察3组治疗1、3和6个月后的NRS评分比较,差异均无统计学意义(P>0.05)。3组治疗12个月后的腰椎(L_(1-4))骨密度比较,差异均有统计学意义(P<0.05),而全髋和股骨颈骨密度比较,差异均无统计学意义(P>0.05),观察1组治疗后的腰椎(L_(1-4))骨密度显著高于观察2组和观察3组,而观察2组和观察3组治疗后的腰椎(L_(1-4))骨密度比较,差异无统计学意义(P>0.05)。3组的不良反应发生率比较,差异无统计学意义(P>0.05)。结论^(99)Tc-MDP治疗绝经后OP的疗效优于唑来膦酸和鲑降钙素,可显著改善患者骨代谢水平和腰椎骨密度,减轻炎症反应,缓解骨痛,且安全性良好。

多模态成像诊断^(99)Tc^(m)-MIBI显像阴性甲状旁腺癌1例

甲状旁腺癌(parathyroid carcinoma,PC)是一种罕见的内分泌肿瘤,相比良性原发性甲状旁腺功能亢进症(primary hyperparathyroidism,PHPT),PC常伴有严重的、难治性高钙血症,更容易诱发高钙危象。PC的预后和总生存期取决于原发肿瘤的大小、肿瘤分期、诊断时的侵袭情况以及早期手术切除所有癌组织的可能性,因而准确的术前影像学评估对于PC治疗至关重要。本文报告了多模态成像协助诊断^(99)Tc^(m)-MIBI显像阴性的PC患者1例,辅助了临床诊疗方案制定。此案例说明,多模态影像可以在PC定性、定位、定量、定期上优势互补,有利于PC的术前诊断及预后评价,有助于临床决策。