Due to the insufficient long-term protection and significant efficacy reduction to new variants of current COVID-19 vaccines, the epidemic prevention and control are still challenging. Here, we employ a capsid and antigen structure engineering (CASE) strategy to manufacture an adeno-associated viral serotype 6-based vaccine (S663V-RBD), which expresses trimeric receptor binding domain (RBD) of spike protein fused with a biological adjuvant RS09. Impressively, the engineered S663V-RBD could rapidly induce a satisfactory RBD-specific IgG titer within 2 weeks and maintain the titer for more than 4 months. Compared to the licensed BBIBP-CorV (Sinopharm, China), a single-dose S663V-RBD induced more endurable and robust immune responses in mice and elicited superior neutralizing antibodies against three typical SARS-CoV-2 pseudoviruses including wild type, C.37 (Lambda) and B.1.617.2 (Delta). More interestingly, the intramuscular injection of S663V-RBD could overcome pre-existing immunity against the capsid. Given its effectiveness, the CASE-based S663V-RBD may provide a new solution for the current and next pandemic.

The rise of nanotechnology has opened new horizons for cancer immunotherapy. However, most nanovaccines fabricated with nanomaterials suffer from carrier-related concerns, including low drug loading capacity, unpredictable metabolism, and potential systemic toxicity, which bring obstacles for their clinical translation. Herein, we developed an antigen self-assembled nanovaccine, which was resulted from a simple acryloyl modification of the antigen to induce self-assembly. Furthermore, a dendritic cell targeting head mannose monomer and a mevalonate pathway inhibitor zoledronic acid (Zol) were integrated or absorbed onto the nanoparticles (denoted as MEAO-Z) to intensify the immune response. The synthesized nanovaccine with a diameter of around 70 nm showed successful lymph node transportation, high dendritic cell internalization, promoted costimulatory molecule expression, and preferable antigen cross-presentation. In virtue of the above superiorities, MEAO-Z induced remarkably higher titers of serum antibody, stronger cytotoxic T lymphocyte immune responses and IFN-γ secretion than free antigen and adjuvants. In vivo, MEAO-Z significantly suppressed EG7-OVA tumor growth and prolonged the survival time of tumor-bearing mice. These results indicated the translation promise of our self-assembled nanovaccine for immune potentiation and cancer immunotherapy.

Objectives:To investigate the clinical and genetic features of young Chinese patients with myeloproliferative neoplasms (MPN) .Methods:In this cross-sectional study, anonymous questionnaires were distributed to patients with MPN patients nationwide. The respondents were divided into 3 groups based on their age at diagnosis: young (≤40 years) , middle-aged (41-60 years) , and elderly (>60 years) . We compared the clinical and genetic characteristics of three groups of MPN patients.Results:1727 assessable questionnaires were collected. There were 453 (26.2%) young respondents with MPNs, including 274 with essential thrombocythemia (ET) , 80 with polycythemia vera (PV) , and 99 with myelofibrosis. Among the young group, 178 (39.3%) were male, and the median age was 31 (18-40) years. In comparison to middle-aged and elderly respondents, young respondents with MPN were more likely to present with a higher proportion of unmarried status (all P<0.001) , a higher education level (all P<0.001) , less comorbidity (ies) , fewer medications (all P<0.001) , and low-risk stratification (all P<0.001) . Younger respondents experienced headache (ET, P<0.001; PV, P=0.007; MF, P=0.001) at diagnosis, had splenomegaly at diagnosis (PV, P<0.001) , and survey (ET, P=0.052; PV, P=0.063) . Younger respondents had fewer thrombotic events at diagnosis (ET, P<0.001; PV, P=0.011) and during the survey (ET, P<0.001; PV, P=0.003) . JAK2 mutations were found in fewer young people (ET, P<0.001; PV, P<0.001; MF, P=0.013) ; however, CALR mutations were found in more young people (ET, P<0.001; MF, P=0.015) . Furthermore, mutations in non-driver genes (ET, P=0.042; PV, P=0.043; MF, P=0.004) and high-molecular risk mutations (ET, P=0.024; PV, P=0.023; MF, P=0.001) were found in fewer young respondents. Conclusion:Compared with middle-aged and elderly patients, young patients with MPN had unique clinical and genetic characteristics.

Background: Granular cell tumor (GCT) is an uncommon tumor, and gastrointestinal tract GCT is even more rare. Aims: To investigate the clinicopathological characteristics, treatment and prognosis of gastrointestinal tract GCT. Methods: Nine cases of gastrointestinal tract GCT from January 2017 to June 2021 at the 903rd Hospital of Joint Logistics Support Force, PLA and Jinhua Municipal Central Hospital Medical Group were retrieved. The clinical data, histopathological characteristics, treatment, and prognosis were retrospectively analyzed. Results: In the 9 patients with gastrointestinal tract GCT, ratio of male to female was 2:1, age at diagnosis was 19-60 years, with a median age of 52 years. Six GCT were found in esophagus, 2 in colorectum and 1 in anus. Endoscopic results showed submucosal protrusion or sessile polyps ranging in size from 2-12 mm with a median of 5 mm. Histology results showed that tumors were located in mucosa and/or submucosa, arranged in solid sheets or nests, with an infiltrative margin and inflammatory infiltrates. Tumor cells were mainly plump and polygonal with abundant cytoplasm and eosinophilic granules. Nuclei were small, the nuclear-cytoplasmic ratio was very low. Mitotic figure was rare. Immunohistochemistry results showed that S100 and CD68 proteins were positive in all patients, SOX10, CD56, Calretinin and Syn were positive in some patients, and CKp, Desmin, SMA, CD117, CD34, Dog1, and α-inhibin were negative in all patients. Esophageal and colorectal GCT patients received endoscopic mucosal resection (EMR) or endoscopic submucosal dissection (ESD). The anal GCT patient underwent local resection. Recurrence or metastasis were not observed during 9-53 months of follow-up. Conclusions: Gastrointestinal tract GCT is rare with non-specific clinical symptoms and submucosal protrusion or sessile polyps under endoscopy. Gastrointestinal tract GCT has special pathomorphology and immunophenotype. EMR or ESD is recommended for small and superficial lesions. Long-term follow-up should be performed.

Objectives:To explore health-related quality of life (HRQoL) and identify its associated variables in Chinese patients with Philadelphia-negative myeloproliferative neoplasms (MPNs) .Methods:In this cross-sectional study, anonymous questionnaires were distributed to adult patients with MPNs to assess symptom burden measured by MPN-10 and HRQoL measured by Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (EORTC QLQ-C30) .Results:The data from 1405 respondents with MPNs, including 645 (45.9%) with essential thrombocythemia (ET) , 297 (21.1%) with polycythemia vera (PV) , and 463 (33.0%) with myelofibrosis (MF) , were analyzed. 646 (46.0%) respondents were male. The median age was 56 (range, 18-99) years. The mean MPN-10 scores were 13.0±12.7, 15.0±14.7, and 21.0±16.6 ( P<0.001) , and the physical component summary (PCS) and mental component summary (MCS) scores were 48.0±8.5, 47.0±9.0, and 42.0±10.0 ( P<0.001) and 51.0±11.0, 50.0±10.8, and 49.0±11.1 ( P=0.002) for respondents with ET, PV, and MF, respectively. Respondents with MF reported the lowest score of physical functioning, role functioning, emotional functioning, cognitive functioning, social function, and global health status (all P<0.01) and the highest score of fatigue, pain, dyspnea, appetite loss, diarrhea, and financial problems (all P<0.05) in EORTC QLQ-C30. Multivariate analyses revealed that higher MPN-10 scores were significantly associated with lower PCS (-0.220 to -0.277, P<0.001) and MCS (-0.244 to -0.329, P<0.001) scores; increasing age (-1.923 to -4.869; all P<0.05) , lower PCS score. Additionally, comorbidity (ies) , symptom at diagnosis, splenomegaly, anemia, unknown driver gene, and higher annual out-of-pocket cost were significantly associated with lower PCS and/or MCS scores. However, age ≥ 60 years, urban household registration, concomitant medication, and receiving ruxolitinib therapy in respondents with MF were associated with higher MCS scores. Weak correlations were found between MPN-10 score (except the subscale of appetite loss and constipation) and EORTC QLQ-C30 score in majority of subscales in respondents with ET (| r| = 0.193-0.457, all P<0.001) , PV (| r| = 0.192-0.529, all P<0.01) , and MF (| r| = 0.180-0.488, all P<0.001) , respectively. Conclusions:HRQoL in patients with MPN was significantly reduced, especially in patients with MF. Sociodemographic and clinical variables were significantly associated with the HRQoL in patients with MPNs.

Objective To analyze oncological outcomes of 125I permanent implant brachytherapy (125I-PIB) in clinically localized prostate cancer. Methods Between June 2008 and June 2015, 121 patients with clinically localized prostate cancer were treated with 125I-PIB, and their average age was 72.37 years old. Before treatment, the average prostate-specific antigen (PSA) level was 17.7 ng/ml, prostate volume was (51.4 ±15.0) cm3, the International Prognostic Scoring System (IPSS) score was (21.6 ±2.4) scores. The intermediate-risk and high-risk patients were treated with adjuvant endocrine or supplemental external beam radiotherapy. All patients were followed-up. Biochemical recurrence was defined as the PSA nadir plus 2.0 ng/ml. Results Average follow-up time of the 121 patients was 41.81 months. The prostate volume was (23.1 ± 10.2) cm3, and the IPSS score was (9.7±3.3) scores. Rates of 5-year overall survival, biochemical recurrence-free survival and cancer-specific survival were 86.7 %, 76.7 % and 96.5 %, respectively. 5-year biochemical recurrence-free survival rates were 88.5 %, 67.5 % and 65.2 % in the low-, intermediate-, and high-risk groups, respectively. Biochemical recurrence-free survival did not differ significantly by three risk groups (P=0.103), but the difference between high-risk and low-risk groups was statistically significant (P=0.028). According to multivariate analysis, higher prostate-specific antigen (P=0.021), higher Gleason score (P=0.023) and higher clinical T stage (P=0.037) were the significant covariates associated with biochemical recurrence-free survival. The addition of hormonal therapy or external beam radiation therapy was associated with significantly better outcomes than brachytherapy monotherapy (P=0.036, P=0.027). As for complications, there were 4 cases of surgery (trans-urethral resection of the prostate) and 4 cases of cardiovascular complications. Conclusions The 125I-PIB can bring excellent oncological outcomes and acceptable complications in patients. Adjuvant endocrine or external beam radiotherapy for the intermediate-risk and high-risk patients may improve the outcome. Factors influencing efficacy include the high PSA, Gleason score and clinical T stage.

In order to meet the needs of national clinical medical education strategy, to train medical students’ability of analyzing and solving problems, it is imminent to carry out the reform on comprehensive experiment in basic medi-cal teaching.During the practice, the basic medical college of Hebei North University tried to integrate functional ex-periment and morphological experiment, and had some experience to share with medical education colleagues.

OBJECTIVETo develop an effective method for isolating and culturing single cancer stem cells.

METHODSThe capillary glass tube was stretched on fire and connected to a sterile plastic tube to prepare the single cell separation apparatus. Single SW480 cell clone spheres in serum-free culture were marked with CD133 and CK7, and the single cancer stem cells were separated and cultivated in 96-well plates or microdrop covered by paraffin.

RESULTSSW480 cell clone formation rate was about 1.04%, and the cell clone spheres highly expressed CD133 with low CK7 expression. The isolation of the single cancer stem cells showed a success rate of 98.99% using the separation device. The cell division profile was comparable between the cell cultures in microdrop and 96-well plates in the initial 2 cell divisions (P>0.05), whereas prolonged cell division occurred afterwards in the microdrop culture as compared to 96-well plate culture. The cell population expansion of the single cancer stem cells was similar between microdrop culture (11.5%, 22/192) and 96-well plate culture (9.2%, 17/184) (P>0.05).

CONCLUSIONSSingle SW480 cells can develop into cancer stem cell spheres. Microdrop culture is convenient and stable, and can be the primary choice for single cancer stem cell culture.

Objective To evaluate the effects of fluid resuscitation and large-volume hemofiltration (HVHF) on the Alveolar-arterial oxygen exchange in patients with refractory septic shock. Method A total of 89 intensive care patients with refractory septic shock treated with fluid resuscitation and/or HVHF were enrolled between August 2006 and December 2009. All the patients were randomly divided into two groups. In group A, patients were treated with fluid resuscitation, n = 41 cases) and in group B, patients were treated with large-volume hemofiltration and fluid resuscitation, n =48). The O2 content of central venous blood(CcvO2), arterial oxygen content (CaO2), Alveolar-arterial oxygen pressure difference (P(A-a)DO2), the ratio of arterial oxygen pressure/alveolar oxygen pressure (PaO2/PAO2), respiratory index (RI) and oxygenation index (OI) were checked. The levels of oxygen exchange in two groups were detected by arterial blood gas analysis before treatnent, 24 hour, 72 hour and 7 days after treatment. The APACHE Ⅱ scores in patients with refractory septic shock were measured before and the 7th day after treatment with HVHF and/or fluid resuscitation respectively. Data were analyzed by using t -test and chi-square test to compare the differences and ratio between two groups and were expressed in mean ± standard deviation, and the analysis of variance was done with SPSS version 12.0 software. Results ① The differences in CcvO2 and CaO2 between two groups were[(0.60±0.24) vs. (0.72±-0.28), P ＜0.05 and (0.84±0.43) vs. (0.94±0.46), P ＜0.05]; and the oxygen extraction rates (O2ER) were significantly different between two groups [(28.7±2.4) vs. (21.7±3.4), P<0.01];② The levels of P(A-a)DO2、ratio of PaO2/PAO2、RI and OI in group B were reduced more significantly than in group A (P<0.05 or P<0.01);③The APACHE Ⅱ scores in both groups were gradually reduced after treatment for 7 days, and the APACHE Ⅱscore in group B on the 7th day of treatment were lower than that in group A[(17.2 ± 6.8) vs. (8.2 ± 3.8), P ＜ 0.01]. Conclusions Fluid resuscitation and HVHF could improve alveolar-arterial-oxygen exchange in patients with refractory septic shock, and at the same time decreased the APACHE Ⅱ scores, improving the survival rate of patients.

Objective To identify the incidence and risk factors of gastrointestinal bleeding in patients supported with mechanical ventilation over 48 hours. Method A total of 127 ICU patients supported with mechanical ventilation for over 48 hours were enrolled from January 1, 2007 to December 31, 2008 for the retrospective study. Exclusion criteria included the history of gastrointestinal bleeding and ulcer, recent gastrointestinal surgery, brain death and active bleeding from nose or throat. Demographics of patients including age, diagnosis at admission, duration of ICU stay, duration of mechanical ventilation, pattern and parameters of ventilation, ICU mortality, A-PACHE II score, multiple organ dysfunction score, and the results of biochemical assays including renal, hepatic and coagulation functions were recorded. Risk factors of gastrointestinal bleeding were analyzed by using univariate analysis And multiple logistic-regression analysis. Results Of the 127 patients, the incidence of gastrointestinal bleeding was 41.7% . and among them 3.9% patients suffered from clinically significant bleeding. However, the independent risk factors of gastrointestinal bleeding were the peak inspiratory pressure > 30 cmH20 (RR = 3.73, 95% CI = 1.59-9.46), renal failure (RR = 1.16,95% CI = 1.02 - 2.32), PLT count <50× 109 L-1(RR = 2.67, 95% CI = 1.32 - 15.78) and prolonged APTT (RR = 4.58, 95%CI = 2.32 - 12.96). The good entetal nutrition had a beneficial effect to the avoidance of gastrointestinal bleeding ( RR = 0.30, 95% CI = 0. 13 - 0.67). Conclusions The incidence of gastrointestinal bleeding is high in patients supported with mechanical ventilation, and the bleeding usually occurs within the first 48 hours. High pressure ventilator setting, renal failure, decreased PLT count and prolonged APTT are significant risk factors of gastrointestinal bleeding. However, the good enteral nutrition is the independent protective factors.