Dynamic tracking analysis of monoclonal antibodies (mAbs) biotransformation in vivo is crucial, as certain modifications could inactivate the protein and reduce drug efficacy. However, a particular challenge (i.e. immune recognition deficiencies) in biotransformation studies may arise when modifications occur at the paratope recognized by the antigen. To address this limitation, a multi-epitope affinity technology utilizing the metal organic framework (MOF)@Au@peptide@aptamer composite material was proposed and developed by simultaneously immobilizing complementarity determining region (CDR) mimotope peptide (HH24) and non-CDR mimotope aptamer (CH1S-6T) onto the surface of MOF@Au nanocomposite. Comparative studies demonstrated that MOF@Au@peptide@aptamer exhibited significantly enhanced enrichment capabilities for trastuzumab variants in comparison to mono-epitope affinity technology. Moreover, the higher deamidation ratio for LC-Asn-30 and isomerization ratio for HC-Asn-55 can only be monitored by the novel bioanalytical platform based on MOF@Au@peptide@aptamer and liquid chromatography-quadrupole time of flight-mass spectrometry (LC-QTOF-MS). Therefore, multi-epitope affinity technology could effectively overcome the biases of traditional affinity materials for key sites modification analysis of mAb. Particularly, the novel bioanalytical platform can be successfully used for the tracking analysis of trastuzumab modifications in different biological fluids. Compared to the spiked phosphate buffer (PB) model, faster modification trends were monitored in the spiked serum and patients' sera due to the catalytic effect of plasma proteins and relevant proteases. Differences in peptide modification levels of trastuzumab in patients' sera were also monitored. In summary, the novel bioanalytical platform based on the multi-epitope affinity technology holds great potentials for in vivo biotransformation analysis of mAb, contributing to improved understanding and paving the way for future research and clinical applications.

Objective:To evaluate the application of metagenomic next-generation sequencing (mNGS) in the diagnosis of osteoarticular infection.Methods:The clinical data of 37 inpatients aged 32-90 year with osteoarticular infection admitted in the Department of Spine Surgery of Qingdao Chest Hospital from January to December 2019 were retrospectively analyzed. There were 31 cases of spine infection and 6 cases of other joint infection. The tissue samples were obtained from the infected sites through puncture or surgical approach in all patients. The tissue samples were subjected to routine culture of mycobacteria, aerobic bacteria and anaerobic bacteria, respectively. The gene amplification and mNGS were performed for detection of mycobacterium tuberculosis DNA (MTB-DNA). The chi-square test or Fisher’s exact test were used to compare the detection rates of pathogen and simple bacterial infection between mNGS and conventional culture. The conventional culture, mNGS and MTB-DNA amplification detection were performed for all samples; with clinical diagnosis as the gold standard, the diagnostic values of 3 methods were evaluated with receiver operating characteristic curve (ROC). Paired sample t test was used to compare white blood cell(WBC) count, erythrocyte sedimentation rate, C-reactive protein of patients before and after treatment. P<0.05 was considered statistically significant. Results:The pathogens were detected by mNGS for 42 times: bacteria for 39 times (92.8%), fungi for twice (4.8%) and Kirks body for once (2.4%). Among 37 patients there were 29 cases of pure bacterial infection (78.4%), 2 cases of pure fungi infection (5.4%), 1 case of pure Kirks body infection (2.7%), and 5 cases of mixed infection of two or more pathogens (13.5%). The detection rates of mNGS and conventional culture were 100.0% (37/37) and 67.6% (25/37), respectively ( χ2=13.987, P<0.05). The detection rates of mNGS and conventional culture in 29 patients with pure bacterial infection were 100.0% (29/29) and 69.0% (20/29), respectively ( χ2=16.913, P<0.05). The area under the ROC curve (AUC) of conventional culture, mNGS, and MTB-DNA in the diagnosis of osteoarticular tuberculosis infection was 0.958 (95% CI: 0.866-1.000, P<0.05), 1.000 (95% CI: 1.000-1.000, P<0.05) and 0.958 (95% CI: 0.866-1.000, P<0.05). All the 37 patients were treated with anti-infective drugs according to the results of mNGS and conventional culture. Among them, 28 patients received surgical intervention. The patients were followed up until April 30, 2020, 1 patient died. After 3 months of follow-up, the WBC count, erythrocyte sedimentation rate and C-reactive protein were (5.5±1.5)×10 9/L, (41±38)mm/h and (5.0±4.6) mg/L, respectively, which were lower than those before anti-infection treatment [(8.0±2.9)×10 9/L, (79±42)mm/h and(63±52)mg/L] ( t=6.536, 8.302 and 6.373, all P<0.05). Conclusion:The metagenomic next-generation sequencing may have important clinical value in the differential diagnosis of osteoarticular infection.

Objective To investigate the correlation between baseline serum lipid levels and hematoma enlargement in patients with acute intracerebral hemorrhage (ICH). Methods From October 2013 to January 2018, patients with ICH admitted to the Department of Neurosurgery, Heze Municipal Hospital, were enrolled retrospectively. The first CT scan was completed within 6 h after onset, and the second one was completed at 48 h after onset. Hematoma enlargement was defined as an increase >33 % in the volume of hematoma on CT. The demographic and baseline clinical data in the hematoma enlargement group and the non -hematoma enlargement group were compared. Multivariate logistic regression analysis was used to identify the independent risk factors for hematoma enlargement. Results A total of 470 patients with acute ICH were enrolled, including 187 females (39.8%) and 283 males (60.2%), aged 47-81 years. Seventy-nine patients (16.8%) had hematoma enlargement. The proportion of patients with atrial fibrillation and who used warfarin before onset, as well as age, baseline National Institutes of Health Stroke Scale score, baseline hematoma volume, international normalized ratio, prothrombin time, activated partial thromboplastin time, and thrombin time of the hematoma enlargement group were significantly higher than those of the non -hematoma enlargement group ( all P< 0.05 ), while from the onset to the first CT scan time, total cholesterol, triglyceride, low-density lipoprotein cholesterol levels were significantly lower than those in the non- hematoma enlargement group (all P<0.05). Multivariate logistic regression analysis showed that baseline total cholesterol <3.20 mmol/L (odds ratio [ OR] 1.32, 95% confidence interval [ CI] 1.08-1.83; P=0.004), baseline hematoma volume≥30 ml (1.76,95% CI 1.30-2.15; P<0.001), and using anticoagulant before onset ( OR 2.37, 95% CI 1.81-3.02; P<0.001 ) had significantly independent correlation with hematoma enlargement. Conclusion Baseline total cholesterol <3.20 mmol/L, hematoma volume ≥30 ml, and using anticoagulant before onset were the independent risk factors for hematoma enlargement in patients with acute ICH.

Objective To observe the clinical effect of auricular bean-burying therapy in acupoints for patients with hepatocellular carcinoma after interventional therapy.Methods A total of 48 patients with hepatocellular carcinoma were divided into control group and observation group.The control group received western conventional nursing.The observation group applied auricular seeds.Pain scores and sleep quality were compared between two groups.Results The intervention group had lower painscores and quality of sleep at 2,3 and 7 d after intervention had lower than the control group (P ＜ 0.05).Conclusion Auricular bean-burying therapy in acupoints can effectively improve postoperative pain,and improve sleep quality for patients with hepatocellular carcinoma after interventional therapy.

Objective To observe the clinical effect of auricular bean-burying therapy in acupoints for patients with hepatocellular carcinoma after interventional therapy.Methods A total of 48 patients with hepatocellular carcinoma were divided into control group and observation group.The control group received western conventional nursing.The observation group applied auricular seeds.Pain scores and sleep quality were compared between two groups.Results The intervention group had lower painscores and quality of sleep at 2,3 and 7 d after intervention had lower than the control group (P ＜ 0.05).Conclusion Auricular bean-burying therapy in acupoints can effectively improve postoperative pain,and improve sleep quality for patients with hepatocellular carcinoma after interventional therapy.

[Objective] The present study aimed to determine the protective role of Melatonin (MT) against Acrylamide (AA)-induced testicular toxicity and the potential molecular mechanism.[Methods] The animals were randomly divided into three groups,the control group (n =12),the AA group (n =12) and the AA+MT group (n =12).The rats in the AA and AA + MT group were gavaged with AA at a dose of 15 mg/(kg · day) for 4 consecutive weeks.After 3 weeks of AA treatment,MT was intraperitoneally injected 30 minutes before AA treatment at 10 mg/(kg· day) for 1 week in the AA + MT rats.Subsequently,the mitochondrial membrane potential measurement,TUNEL assay,Western blot and electron microscopic techniques were applied in the present study.[Results] The results showed that AA could decrease the testis mitochondrial membrane potential (P ＜ 0.05) which could be recovered by MT (P ＜ 0.05).Moreover,MT induced down-regulation of Bax expression and up-regulation of Bcl-2 expression in the testis,compared with AA rats.The amelioration of testicular apoptosis was further confirmed by the TUNEL labeling.Western blot results suggested that the decreased ratios of Bcl-2/Bax and Bcl-xL/Bak in the AA group (both P ＜ 0.05) could be recovered by MT treatment (both P ＜ 0.05).The levels of Cyt-c,Casp-3,p53 and NF-κB in AA group were markedly elevated compared with the control (all P ＜ 0.05),and reduced in MT treatment group (all P ＜ 0.05).MT could relieve abnormal mitochondrial structures in the seminiferous tubule in the electron microscopic level.[Conclusion] MT may exert productive effect through its anti-apoptotic properties associated with mitochondria.

UNLABELLEDTo study the effect of a Chinese herbal prescription on external calcium deposition to weight-bearing bone in simulated weightlessness rats.

METHODTwenty-one male Wistar rats were divided into 3 groups: control group, tail suspension group, tail suspension with Chinese medicine group which takes a Chinese herbal prescription extract (containing Radix Rehmanniae Preparata, Radix Acanthopanacis Bidentatae, Radix Astragali, Radix Angelicae Sinensis, Concha Ostreae prepared by acetic acid) by intragastric administration. After 1 week adaption, there start off 3 weeks simulated weightlessness by tail suspension. At the eleventh day of simulated weightlessness, every rat was given one equal dose of 41Ca tracer by intragastric administration. Right femurs were separated as experiment over, and the ratio of 41Ca to 40Ca (41Ca/40Ca) was measured by accelerator mass spectrometry (AMS), while total femur calcium was measured by inductively coupled plasma-atomic emission spectroscopy (ICP-AES). Femur 41Ca deposition amount (DA) and femur 41Ca deposition ratio (DR) were calculated.

RESULTThe results showed that compared with control group, 41Ca/40Ca decreased significantly (P < 0.001) in tail suspension group, while in tail suspension with Chinese medicine group, it significantly increased (P < 0.05). DA and DR were both decreased significantly (P < 0.001) in tail suspension group, but no significant change in tail suspension with Chinese medicine group as compared with control group. Compared with tail suspension group, DA and DR increased significantly (P < 0.001) in tail suspension with Chinese medicine group.

CONCLUSIONSimulated weightlessness by tail suspension can cause decreased deposition of external calcium to weight-bearing bone, and the Chinese herbal prescription in this trial is effective to prevent the decrease. Moreover, multiple mechanisms may contribute to weightlessness induced osteoporosis, besides calcium deposition disturbance.

Animals ; Bone Resorption ; etiology ; metabolism ; Calcification, Physiologic ; drug effects ; Calcium ; metabolism ; Calcium Radioisotopes ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Femur ; drug effects ; metabolism ; Hindlimb Suspension ; Male ; Mass Spectrometry ; Rats ; Rats, Wistar ; Weightlessness Simulation ; adverse effects

Objective To study the effect of Chinese herb medicine compound on general state and bone-muscle system in simulated weightlessness rats, and to observe the synergistic action of other ingredients in the compound on calcium. Methods Thirty Wistar rats were divided into 3 groups:control group, tail suspend group, tail suspend and medicine group which were given Chinese herb medicine compound by intragastric administration. After 3 weeks simulated weightlessness, body weight (BW), muscle weight (MW) and index (MI) of posterior limb, bone length (BL), wet weight (BWW), index (BI), dry weight (BDW), content of organic (ORG) and inorganic (INO) substance, bone mineral density (BMD), and mechanical properties (MEC) of femur were observed. Results At the middle-later stage of the experiment, BW of tail suspend group decreased significantly (P

Objective To investigate the effect of hindlimb unloaded 3 weeks on bone mineral density of different parts in simulated weightlessness rats,and the effect of Chinese herbal prescriptions on them.Methods 2-month-old male Wistar rats were randomly divided into 3 groups:control group(CON),hindlimb unloading group(HU),HU+Chinese herbal prescription group(HUC).During the whole experiment(4 weeks),HUC were given Chinese herbal prescriptions(contains Radix Acanthopanacis Senticosl,Radix Rehmanniae Praeparata,Radix Achyranthis Bidentatae,Concha Ostreae,etc),while CON and HU were given equal measures Concha Ostreae prepared by acetic acid.After 1 week of adaption period,weightlessness was simulated by tail suspension in HU and HUC for three weeks.Rats were executed at the end of the fourth week and bone mineral density of 7 differernt parts were determined.BMD of skull bone and second thoracic vertebra were examined by buoyancy method,and fourth lumbar vertebra,humerus,femur,radio-ulna and tibia were examined by bone densitometer.Results Compared with CON,BMD of skull of HU increased significantly(P 0.05),but BMD of lumbar vertebra,femur and tibia were decreased notably(P 0.05).Conclusions The change of hindlimb unloaded 3 weeks on systemic BMD presented certain trend.BMD of skull bone increased,little change on upper limbs and second thoracic vertebra,and fourth lumbar vertebra and hind limbs reduced.Chinese herbal prescriptions had the function to relieve weightlessness osteoporosis,and had positive effect on calcium distribution with weightlessness.

PURPOSE : To evaluate clinical and three dimensional (3D) dosimetric parameters associated with esophageal injury after radiotherapy for non-small cell lung cancer (NSCLC). MATERIALS AND METHODS : The records of 254 patients treated for NSCLC between 1992 and 2001 were reviewed. A variety of metrics describing the esophageal dose were extracted. Chemotherapy was given in 143 patients (56%). The RTOG toxicity criteria for grading of esophageal injury were used. The median follow-up time of all patients was 43 months with the range of 0.5~120 months. Logistic regression, contingency table analyses and Fisher's exact tests were used for statistical analysis. RESULTS : Acute toxicity occurred in 78% patients (199/254); grade 1,138; grade 2, 38; grade 3, 22 and grade 4, 1. For acute toxicity> or =Grade 2, BID-RT, age, nodal stage> or =N2, and most dosimetric parameters were predictive. Late toxicity occurred in 17 (7%) of 238 patients; grade 1, 5; grade 2, 4; grade 3, 5 and grade 4, 3. The median and maximum time to onset of late toxicity was 5 and 40 months after radiotherapy, respectively. Late toxicity occurred in 2%, 3%, 17%, 26%, and 100% of patients with acute grade 0, 1, 2, 3 and 4 toxicity, respectively. For late toxicity, the severity of acute toxicity was most predictive. CONCLUSION : A variety of dosimetric parameters are predictive for acute and late esophageal injury. A strong correlation between the dosimetric parameters prevented a comparison between the predictive abilities of these metrics. The presence of acute injury was the most predictive factor for the development of late injury. This suggests that late injury may be "consequential" and that aggressive treatment of acute effects may reduce the risk of late injury. Additional studies to better define predictors of RT-induced esophageal injury are needed
Carcinoma, Non-Small-Cell Lung ; Drug Therapy ; Follow-Up Studies ; Humans ; Logistic Models ; Radiotherapy