China ; Humans ; Nanostructures ; toxicity ; Nanotechnology

IL-18, as a polyphonic cytokine, is important in immune response and physiologic function. We designed one pair of primers, amplified the porcine IL-18 gene fused with a C-terminal 6xHistidine tag, and then subcloned into the pFastBacDual of Baculovirus transfer vector and transformed into DH10Bac containing a shuttle vector of Bacmid. After co-transfecting the recombinant plasmid into insect cells, the 18 kDa expressed protein of porcine IL-18 was detected by SDS-PAGE; the specificity of expressed protein was confirmed by Western blotting. The purified porcine IL-18 protein induced obvious proliferation of porcine T lymphocytes in vitro, which indicated that the expression of IL-18 had high biological activity.
Animals ; Baculoviridae ; genetics ; metabolism ; Cells, Cultured ; Genetic Vectors ; genetics ; Insecta ; genetics ; metabolism ; Interleukin-18 ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Swine ; Transfection

OBJECTIVETo develop a quartz crystal microbalance (QCM) immunosensor with high sensitivity and selectivity for the rapid detection of diethylstilbestrol.

METHODSDextran was used as reducing agent for preparing gold nanoparticles (AuNPs) with the size of 40 nm. The AuNPs were coupled with anti-DES antibody after amination. A monolayer was generated after immersing the quartz crystal into the solution of 5 mmol/L 11-mercaptoundecanoic acid(MUA) for 16 hours. After the monolayer was activated by 1-ethyl-3-(3-dimethylaminopropry) carbodiimide hydrochloride (EDC·HCl) and N-hydrosuccinimide (NHS), 20 μl of 2.2 mg/ml DES-HS-BSA was dropped onto the surface of crystal to prepare a sensitive membrane which can recognize DES specifically. Then, 50 μl of 1 mol/L ethanolamine (pH 8.5) was used to seal the carboxylic groups to make the sensitive membrane which could identify DES specifically. QCM immunosensor was used as detection platform to optimize the reaction conditions. Under the optimized conditions, 10 μl of 28 μg/ml AuNPs-antibody was mixed with 10 μl of 0.03-2.5 μg/ml DES, and the mixture was added on the sensitive membrane. QCM immunosensor was used to detect the signals and the standard curve was obtained at the same time. The detection limit was calculated based on the standard curve. The specificity was evaluated by testing DES and its analogues with the same concentration.

RESULTSThe optimized concentration for the immobilization of DES-HS-BSA on the surface of QCM was 2.2 mg/ml. The optimized concentration for coupling anti-DES antibody with AuNPs was 7 μg/ml and 15 nmol/L, respectively. The optimized concentration of AuNPs-antibody was 14 μg/ml. The logarithm of DES concentration was proportional to the frequency shift in the range of 0.16-500 ng/ml, Δf=-24.170 lgCDES+69.71, R(2)=0.998. The detection limit of this method was 0.13 ng/ml. DES analogues could not influence the detection of DES obviously, so the sensor had good specificity.

CONCLUSIONThe quartz crystal microbalance immunosensor with gold nanoparticals amplification could detect DES sensitively and rapidly.

Renal cell carcinoma is one of the common malignant tumors in genitourinary cancer.20％-30％ of patients have distant metastases.However,current biomarkers used for diagnosis,recurrence monitoring,and prognosis assessment are still uncertain.Circulating tumor cells (CTCs) are tumor cells that are detached from the primary tumor or metastases,and invade and presenting in peripheral blood vessels.Therefore,CTCs is regarded as a key step in the process of tumor metastasis.However,current studies on CTCs of renal cell carcinoma are facing with problems such as inconsistent detection methods and limited clinical applications.This article reviews the progress in the detection and clinical application of CTCs of renal cell carcinoma.

Objective:To analyze the biochemical failure rate and its predictive factors after radical prostatectomy (RP) in patients with high-risk localized prostate cancer.Methods:The data of 166 patients with high-risk localized prostate cancer who underwent RP surgery in Peking university cancer hospital from January 2015 to November 2021 were retrospectively reviewed. The average age was 65.4±6.2 years old, and the average body mass index (BMI) was 24.86±3.23 kg/m 2. The median prostate-specific antigen (PSA) was 19.84 (10.98, 44.47) ng/ml, PSA density was 0.68 (0.34, 1.32)ng/ml 2, and prostate volume was 31.20 (25.58, 40.23) ml. Biopsy pathology Gleason score according to the International society of Urological Pathology(ISUP) grade group: 18 cases of group 1, 33 cases of group 2, 30 cases of group 3, 51 cases of group 4, and 33 cases of group 5, 1 case was unknown. The percentage of puncture positive needles was (55.4±25.7)%, and the largest linear length of positive lesions was 80.0% (60.0%, 90.0%). Preoperative clinical stage : 14 cases in ≤T 2b stage, 117 cases in T 2c stage, 13 cases in T 3a stage and 22 cases in ≥T 3b stage; 157 cases in N 0 stage, 9 cases in N 1 stage. One hundred and three patients (62.0%) were assessed by traditional imaging and 63(38.0%) were assessed by PSMA PET-CT. The patients underwent laparoscopic radical prostatectomy. 64 patients (38.6%) received neoadjuvant therapy, including 37 received neoadjuvant therapy for 1-3 months, 23 for 4-6 months and 4 for over 6 months. The postoperative pathological characteristics, treatment and prognosis of the patients were analyzed. The primary endpoint was biochemical failure, including biochemical persistence(BCP, defined as PSA≥0.1ng/ml at 4-6 weeks after operation, and confirmed by re-examination at least 1 week interval) and biochemical recurrence(BCR, PSA falling below 0.1ng/ml after operation and then rising ≥0.2 ng/ml without adjuvant therapy or after the end of adjuvant treatment). Results:Compared with preoperative clinicopathological characteristics, 48(28.9%) cases had postoperative pathological ISUP upgrade, 98 (59.0%)cases had T stage upgrade, and 13 (7.8%) cases had N stage upgrade. The rate of positive margins was 53%, and apex margin was the most common positive site (65.9%). The postoperative PSA in 114 patients (68.7%) decreased to less than 0.1ng/ml, of which 74 patients didn't receive the therapy and 40 patients received adjuvant therapy. 52 patients (31.3%) had postoperative PSA more than 0.1ng/ml and among them, 51 cases received salvage treatment. 5 patients (3.0%) underwent PSA progression during adjuvant or salvage endocrine therapy and were considered to have castration resistance. After a median follow-up time of 25.5 (12.0, 40.0) months, 78 patients (48.4%, 78/161) experienced biochemical failure, including 49 BCP and 29 BCR, the median time of biochemical failure was 30.0 (95% CI 14.5-45.5) months. Adjuvant therapy could reduce the rate of BCR (31.1% and 15.8%, P=0.08). Baseline PSA, PSA density, proportion of pathological ISUP ≥4, proportion of pathological T stage ≥T 3a, adjuvant therapy, and positive surgical margins were significantly associated with biochemical failure ( P=0.034, 0.002, 0.004, 0.025, <0.001and 0.047). Multivariate Cox regression analysis showed that adjuvant therapy ( P<0.001, OR=0.12), PSA density ( P=0.03, OR=1.19) and positive surgical margins ( P=0.034, OR=1.80) were independent factors for biochemical failure. Conclusions:Patients with high-risk localized prostate cancer have a high rate of biochemical failure after RP and need to receive RP-based multimodal therapy. Adjuvant therapy, PSA density and positive surgical margins are independent factors associated with postoperative biochemical failure.

Objective:To explore the safety and efficacy of laparoscopic microwave ablation combined with partial nephrectomy in the treatment of cystic renal masses.Methods:The 19 patients with cystic renal masses undergoing laparoscopic microwave ablation combined with partial nephrectomy from November 2017 to December 2019 were retrospectively analyzed. There were 13 males and 6 females. The average age was 46.2 years. The mean body mass index was (25.8±3.1) kg/m 2. The masses located in the left kidney in 7 cases and the right kidney in 12 cases. The ECOG scores were 0. The mean maximum diameter of the tumors was (2.8±1.3) cm. Five cases were diagnosed with Bosniak Ⅲ and 14 cases with Bosniak Ⅳ. According to R. E.N.A.L. scoring, 11 cases were of low difficulty (4-6 points), 7 cases of medium difficulty (7-9 points) and 1 case of high difficulty (10-12 points). The cystic renal masses were ablated by laparoscopic microwave ablation, then followed by partial nephrectomy. Postoperative complications were observed and the prognosis was assessed by CT or MRI. Results:The mean duration of operation was (84.0±20.8) min. The median intraoperative blood loss was 20 (10-50) ml. The median duration of postoperative hospitalization was 3 (2-6) d, and no complications such as bleeding, infection, gross hematuria or urine leakage were observed. According to the malignant degree of cystic renal masses, the patients were divided into low-risk group and high-risk group. The patients with benign cystic kidney tumors or with low biological malignancy were considered as the low-risk group, while the patients with high malignant pathology were considered as the high-risk group. In the low-risk group, there were 4 patients, including 1 patient with papillary adenoma, 1 patient with renal angiomyolipoma, 1 patient with low-grade malignant potential multilocular cystic renal tumor, and 1 patient with renal chromophobe carcinoma (stage T 1a). In the high-risk group, there were 15 cases, including 14 cases of clear cell renal cell carcinoma (AJCC pathological stage: T 1a stage 11 cases, T 1b stage 3 cases; WHO/ISUP classification: 7 cases in grade 1, 6 cases in grade 2, and 1 case in grade 3); 1 case of Type 2 papillary renal cell carcinoma (stage T 1b, grade 2). The median follow-up was 20 months (12-37 months). Both groups survived, and no signs of tumor recurrence, implantation or metastasis were found in chest and abdomen imaging. Blood tests were performed regularly, and no significant abnormalities occurred. Conclusions:The safety and efficacy of laparoscopic microwave ablation combined with partial nephrectomy for the treatment of cystic renal masses is satisfactory, and postoperative pathology is clear, providing a potential option for cystic renal masses treatment.

Objective: To investigate the relationship between SUVmax on preoperative 68Ga-PSMA PET-CT and the clinicopathological characteristics of patients treated with radical prostatectomy. Methods: The clinicopahtological data of patients evaluated with ⁶⁸Ga-PSMA PET-CT preoperatively and treated with radical prostatectomy between May 2016 and August 2019 were retrospectively reviewed. 31 patients with a mean age (63.1±4.9) and baseline PSA (72.71±173.15)ng/ml were enrolled. Their BMI mean (24.6±3.0)kg/m². Baseline testosterone of 14 patients was (4.72±1.64)ng/ml.Based on the Gleason scores related ISUP classification, all patients were classified into grade one in 5 cases, grade 2in 7 cases, grade 3 in 4 cases, grade 4 in 10 cases and grade 5 in 5 cases. The clinical classification included 6 cases in T_2a stage, 2 cases in T_2b stage, 17 cases in T_2c stage, 1 case in T_3a stage, 4 cases in T_3b stage and 1 case in T₄ stage. SUVmax was accessed by two independent professional nuclear medicine physicians. SUVmax was 12.49±9.38. SPSS 16.0 software was used to do statistic analysis. Results: The post-operative pathological results showed the surgical margin positive in 19 cases, negative in 12 cases, vascular positive in 5 cases, negative in 20 case, positive nerve invasion in 20 cases and negative in 11 cases. 2 patients were low risk, 7 patients were medium risk and 22 patients were high risk according to D′Amico classification. Based on the basis of PSA(≤10 or>10) and Gleason score(≤6 or>6), 6 patients were in group with low PSA and low Gleason score, 5 patients were low PSA and high Gleason score, 9 patients were high PSA and low Gleason score, 11 patients were high PSA and high Gleason score. SUVmax had a significant positive relationship with pathological ISUP(r=0.434, P=0.015) and SUVmax in patients with positive intravascular tumor emboli was significantly higher than those with negative intravascular tumor emboli(14.78±10.68 vs. 8.17±2.81, P=0.005). No significant correlation was found between SUVmax and baseline PSA, testosterone, pathologic T stage, surgical margin, nerve invasion, pelvic lymph node status as well as risk stratification. SUVmax could distinguish pathologic ISUP grade 5 with a maximum AUC 0.747 (P=0.033) and the sensitivity was 88.9%. The specificity was 77.3% when SUVmax≥11.34. SUVmax in patients with upgrading ISUP was significantly higher than that in patients with downgrading ISUP (16.01±5.40 vs. 4.98±2.11, P=0.007). Conclusions SUVmax measured on preoperative ⁶⁸Ga-PSMA PET-CT may have a clinical significance in predicting unfavorable pathological factors for patients treated with radical prostatectomy.

Objective: To analyze and compare therapy responses, outcomes, and incidence of severe hematologic adverse events of flumatinib and imatinib in patients newly diagnosed with chronic phase chronic myeloid leukemia (CML) . Methods: Data of patients with chronic phase CML diagnosed between January 2006 and November 2022 from 76 centers, aged ≥18 years, and received initial flumatinib or imatinib therapy within 6 months after diagnosis in China were retrospectively interrogated. Propensity score matching (PSM) analysis was performed to reduce the bias of the initial TKI selection, and the therapy responses and outcomes of patients receiving initial flumatinib or imatinib therapy were compared. Results: A total of 4 833 adult patients with CML receiving initial imatinib (n=4 380) or flumatinib (n=453) therapy were included in the study. In the imatinib cohort, the median follow-up time was 54 [interquartile range (IQR), 31-85] months, and the 7-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.2%, 88.4%, 78.3%, and 63.0%, respectively. The 7-year FFS, PFS, and OS rates were 71.8%, 93.0%, and 96.9%, respectively. With the median follow-up of 18 (IQR, 13-25) months in the flumatinib cohort, the 2-year cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) were 95.4%, 86.5%, 58.4%, and 46.6%, respectively. The 2-year FFS, PFS, and OS rates were 80.1%, 95.0%, and 99.5%, respectively. The PSM analysis indicated that patients receiving initial flumatinib therapy had significantly higher cumulative incidences of CCyR, MMR, MR(4), and MR(4.5) and higher probabilities of FFS than those receiving the initial imatinib therapy (all P<0.001), whereas the PFS (P=0.230) and OS (P=0.268) were comparable between the two cohorts. The incidence of severe hematologic adverse events (grade≥Ⅲ) was comparable in the two cohorts. Conclusion: Patients receiving initial flumatinib therapy had higher cumulative incidences of therapy responses and higher probability of FFS than those receiving initial imatinib therapy, whereas the incidence of severe hematologic adverse events was comparable between the two cohorts.
Adult ; Humans ; Adolescent ; Imatinib Mesylate/adverse effects* ; Incidence ; Antineoplastic Agents/adverse effects* ; Retrospective Studies ; Pyrimidines/adverse effects* ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy* ; Treatment Outcome ; Benzamides/adverse effects* ; Leukemia, Myeloid, Chronic-Phase/drug therapy* ; Aminopyridines/therapeutic use* ; Protein Kinase Inhibitors/therapeutic use*

Background: Cardiopulmonary resuscitation (CPR) strategies in COVID-19 patients differ from those in patients suffering from cardiogenic cardiac arrest. During CPR, both healthcare and non-healthcare workers who provide resuscitation are at risk of infection. The Working Group for Expert Consensus on Prevention and Cardiopulmonary Resuscitation for Cardiac Arrest in COVID-19 has developed this Chinese Expert Consensus to guide clinical practice of CPR in COVID-19 patients. Main recommendations: 1) A medical team should be assigned to evaluate severe and critical COVID-19 for early monitoring of cardiac-arrest warning signs. 2) Psychological counseling and treatment are highly recommended, since sympathetic and vagal abnormalities induced by psychological stress from the COVID-19 pandemic can induce cardiac arrest. 3) Healthcare workers should wear personal protective equipment (PPE). 4) Mouth-to-mouth ventilation should be avoided on patients suspected of having or diagnosed with COVID-19. 5) Hands-only chest compression and mechanical chest compression are recommended. 6) Tracheal-intubation procedures should be optimized and tracheal-intubation strategies should be implemented early. 7) CPR should be provided for 20-30 min. 8) Various factors should be taken into consideration such as the interests of patients and family members, ethics, transmission risks, and laws and regulations governing infectious disease control. Changes in management: The following changes or modifications to CPR strategy in COVID-19 patients are proposed: 1) Healthcare workers should wear PPE. 2) Hands-only chest compression and mechanical chest compression can be implemented to reduce or avoid the spread of viruses by aerosols. 3) Both the benefits to patients and the risk of infection should be considered. 4) Hhealthcare workers should be fully aware of and trained in CPR strategies and procedures specifically for patients with COVID-19.