Objective To construct and identify norepinephrine ( NE) complete antigen for the preparation of high sensitive and specific anti-NE monoclonal antibody .Methods Glutaraldehyde ( GA) and 1-Ethyl-3-(3-Dimethylaminopropyl ) carbodiimide ( EDC) were used to cross-link NE with carrier pro-teins (BSA, OVA) for NE complete antigen preparation under conditions of pH 4.5 or pH9.0.Three assays including UV scanning , SDS-PAGE and FeCl3 color reaction were performed for identification of NE com-plete antigen.Serum antibody titers were evaluated in mice model induced by intraperitoneal immunization with NE complete antigen .Results NE complete antigens were successfully prepared as indicated by the three identification assays .The coupling ratio was significantly increased in a time-depended manner under the condition of pH9.0 in comparison to that in the condition of pH 4.5.Indirect ELISA results showed that , when coating antigens and serum antibodies were prepared with the same cross -linking method , the serum antibody titers were significantly higher than those with different methods .Conclusion Anti-NE antibodies were successfully prepared by immunizing mice with NE complete antigens .

Objective To evaluate the safety and efficacy of induction chemotherapy with gemcitabine followed by concurrent chemoradiotherapywith capecitabine in patients with locally advanced pancreatic cancer (LAPC).Methods A total of 42 patients with locally advanced pancreatic cancer were enrolled.All patients received seven cycles of induction chemotherapy of gemcitabine 1 000 mg/m2,once a week.Concurrent chemoradiotherapy began 1 week after completion of induction chemotherapy.Radiotherapy was delivered with a median dose of 54 Gy (34-64 Gy) with 1.8-2.0 Gy in a fraction.The radiotherapy was combined with capecitabin at a dosage of 825 mg/m2 twice daily,5 d/week.Results Twenty patients (47.6％) were evaluated as clinical benefit response (CBR).Two cases were observed with complete remission (CR),8 with partial remission (PR),27 with stable disease (SD),and 5 with progressive disease (PD).The median overall survival was 10.1 months (range of 4-36 months).The 1-,2-year overall survival rate was 38.2％ and 18.2％,respectively.Myelosuppression was recorded in 20 patients with grades 1-2,and 5 patients with grade 3.Twenty-two patients suffered from grade 1-2 gastrointestinal toxicities,while 4 patients suffered from grade 3.Conclusions The preliminary results showed that induction chemotherapy with gemcitabine followed by concurrent chemoradiotherapy with capecitabine in patients with LAPC might achieve encouraging efficacy with better tolerance.

Objective To study the feasibility of non-invasive spatially-resolved near-infrared spectroscopy (SR-NIRS) in clinical bedside monitoring of shock.Methods The central venous blood samples of 25 patients with shock were collected and the central internal jugular central vein oxygenation (ScvO2) level was measured.The self-developed non-invasive SR-NIRS device was used to measure tissue blood oxygen saturation (StO2) surrounding the region of jugular central vein.In addition,the artery oxygen saturation (SaO2) and partial pressure of oxygen (PO2) were also measured using conventional methods.The correlation between StO2 between ScvO2,SaO2 and PO2 was analyzed.Results StO2 levels in shock patients were highly correlated with ScvO2 levels (r=0.84,P＜0.001) and the concordance coefficient of 0.80 was high.Conclusion StO2 value collected from the surrounding region of jugular central vein by SR-NIRS device can be used as an indicator of shock suggesting the potential of noninvasive SR-NIRS for bedside shock monitoring.

A fluorescent microsphere-based immunochromatographic strip test (FICT) was developed for the rapid, sensitive, and quantitative detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibodies at the pen-side. The assay was based on the formation of a sandwich immune-complex (anti-pig IgG-PRRSV antibodies-NSP7/N), which was validated by a comparison with IDEXX-ELISA using 3325 clinical specimens. The diagnostic specificity, sensitivity, and accuracy of FICT were 97.28, 93.41, and 94.95%, respectively. FICT showed a good correlation with the virus neutralization assay. Overall, a promising pen-side diagnostic tool was developed for the rapid and quantitative detection of PRRSV antibodies within 15 min.

Objective: To investigate the expressions of Fra-1 and HMGA1 in laryngeal squamous cell carcinoma and their correlation. Methods: Immunohistochemistry and reverse transcription-polymer chain reaction (RT-PCR) were used to detect the expressions of HMGA1 and Fra-1 in laryngeal squamous carcinoma tissues in 47 cases and para-carcinoma tissues in 21 cases(the First Hospital of Shijiazhuang). The relationship between the gene expressions in carcinoma tissues and clinopathological parameters such as pathological grade, clinical stage, lymph metastasis, age and anatomic site and the relevance of the two gene expressions were analyzed. SPSS 13.0 software was used to analyze the data. Results: The positive expression rates of Fra-1 and HMGA1 proteins in laryngeal squamous cancer tissue were 48.9% and 53.2%, which were respectively higher than the rates of 19.0% for Fra-1 (χ²=5.416, P<0.05) and of 23.8% for HMGA1 (χ²=5.083, P<0.05) in adjacent tissues. The expression of Fra-1 gene was correlation with pathological grade, clinical stage and lymph metastasis (t values were -1.079, -1.066 and -1.067, all P<0.05), but not with age and anatomic site (t values were -1.068 and -1.054, both P>0.05). The expression of HMGA1 gene was correlation with pathological grade, clinical stage, lymph metastasis and age (t values were -1.112, -1.065, -1.009 and -1.066, all P<0.05), but not with anatomic site (t=-1.036, P>0.05). The expressions of Fra-1 and HMGA1 gene were positively correlation (r=0.672, P<0.05). Conclusions In laryngeal squamous cancer, Fra-1 and HMGA1 are excessive expression, with a positive correlation between the expressions of both genes.

Objective: To study the expression of miR-142-5p in lung adenocarcinoma tissues, and to explore its effect on proliferation, invasion, migration and epithelieal-mesenchymal transition (EMT) of H1650 cells and the potential mechanisms. Methods:Atotal of 107 pairs of lung adenocarcinoma tissues and corresponding para-cancerous tissues from patients, who underwent tumor resection and were pathologically confirmed at the Department of Thoracic Surgery, the Fourth Hospital of Hebei Medical University between Jan. 2014 and Jan. 2015, were collected for this study; in addition, human lung adenocarcinoma cell lines (H1650, HCC827, A549, H1975, PC9) and human bronchial epithelial BEAS-2B cells were also used in this study. qPCR was used to detect the expression of miR-142-5p in lung adenocarcinoma tissues and cell lines. The correlation between expression of miR-142-5p and clinical features was analyzed.After transfection with miR-142-5p mimics or miR-negative control (miR-NC) plasmid, the proliferation, invasion and migration of H1650 cells were detected with CCK-8, Transwell invasion assay and Wound healing assay, respectively. The bioinforamtics tool was used to predict the target genes of miR-142-5p, and Luciferase reporter gene assay was performed to validate the regulation of miR-142-5p on target gene. Western blotting (WB) was used to detect the expressions of cyclin-dependent kinase 5 (CDK5) and EMTrelated protein. Results: Compared to Para-cancerous tissues and BEAS-2B cells, the expression of miR-142-5p was lower in lung adenocarcinoma tissues and cell lines (all P<0.01). Of the 107 cases of lung adenocarcinoma tissues, 61 cases (57.01%) showed decreased miR-142-5 expression, which was correlated with the TNM stage and lymph node metastasis (both P<0.01). Transfection of miR-142-5p mimics significantly up-regulated the expression of miR-142-5p and decreased the proliferation, invasion and migration of H1650 cells (all P<0.05 or P<0.01). Bioinformatics showed that CDK5 was a target gene of miR-142-5p. Luciferase reporter gene assay and WB validated that miR-142-5p could significantly down-regulate CDK5 expression in H1650 cells, up-regulate the expression of E-cadherin and down-regulate the expressions of N-cadherin, Twist and Snail in H1650 cells (all P<0.01). Conclusion: miR-142-5p is low expressed in lung adenocarcinoma tissues and cell lines; it suppresses the EMT process to inhibit, invasion and migration of H1650 cells via down-regulating the expression of CDK5.

Background::T-cell-mediated immunity is crucial for the effective clearance of viral infection, but the T-cell-mediated immune responses that are induced by booster doses of inactivated severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines in people living with human immunodeficiency virus (PLWH) remain unclear.Methods::Forty-five PLWH who had received antiretroviral therapy (ART) for more than two years and 29 healthy controls (HCs) at Beijing Youan Hospital were enrolled to assess the dynamic changes in T-cell responses between the day before the third vaccine dose (week 0) and 4 or 12 weeks (week 4 or week 12) after receiving the third dose of inactivated SARS-CoV-2 vaccine. Flow cytometry, enzyme-linked immunospot (ELISpot), and multiplex cytokines profiling were used to assess T-cell responses at the three timepoints in this study.Results::The results of the ELISpot and activation-induced marker (AIM) assays showed that SARS-CoV-2-specific T-cell responses were increased in both PLWH and HCs after the third dose of the inactivated SARS-CoV-2 vaccine, and a similar magnitude of immune response was induced against the Omicron (B.1.1.529) variant compared to the wild-type strain. In detail, spike-specific T-cell responses (measured by the ELISpot assay for interferon γ [IFN-γ] release) in both PLWH and HCs significantly increased in week 4, and the spike-specific T-cell responses in HCs were significantly stronger than those in PLWH 4 weeks after the third vaccination. In the AIM assay, spike-specific CD4 + T-cell responses peaked in both PLWH and HCs in week 12. Additionally, significantly higher spike-specific CD8 + T-cell responses were induced in PLWH than in HCs in week 12. In PLWH, the release of the cytokines interleukin-2 (IL-2), tumour necrosis factor-alpha (TNF-α), and IL-22 by peripheral blood mononuclear cells (PBMCs) that were stimulated with spike peptides increased in week 12. In addition, the levels of IL-4 and IL-5 were higher in PLWH than in HCs in week 12. Interestingly, the magnitude of SARS-CoV-2-specific T-cell responses in PLWH was negatively associated with the extent of CD8 + T-cell activation and exhaustion. In addition, positive correlations were observed between the magnitude of spike-specific T-cell responses (determined by measuring IFN-γ release by ELISpot) and the amounts of IL-4, IL-5, IL-2 and IL-17F. Conclusions::Our findings suggested that SARS-CoV-2-specific T-cell responses could be enhanced by the booster dose of inactivated COVID-19 vaccines and further illustrate the importance of additional vaccination for PLWH.

[Abstract] Objective: To investigate the effect of long non-coding RNA DiGeorge syndrome critical region gene 5 (DGCR5) on proliferation, invasion and migration of esophageal squamous cell carcinoma (ESCC) TE1 cells and its mechanism. Methods: qPCR was used to detect the expression level of DGCR5 in ESCC cell lines (TE1, Yes-2, KYSE150 and Eca9706). TE1 cells were transfected with siRNA-DGCR5(si-DGCR5) and negative control (si-NC) plasmids, respectively. CCK-8, Wound healing and Transwell assay were used to detect the proliferation, migration and invasion of TE1 cells before and after DGCR5 knockdown. The relationship between DGCR5 expression and epidermal growth factor receptor (EGFR) in ESCC tissues was analyzed by GEPIA database. The mRNA and protein expressions of EGFR in ESCC cell line were examined by qPCR and Western blotting (WB). WB was further used to detect the expression of EGFR protein in TE1 cells before and after DGCR5 knockdown. Results: lncRNA DGCR5 was highly expressed in ESCC cell lines (all P<0.01). qPCR confirmed that the expression of DGCR5 in TE1 cells of si-DGCR5 group was significantly lower than that of si-NC group (P<0.01). The proliferation, migration and invasion ability of TE1 cells in si-DGCR5 group were significantly lower than those in si-NC group (all P<0.01). GEPIA database showed that the expression of DGCR5 was positively correlated with EGFR in ESCC tissues (P<0.01). WB showed that the protein level of EGFR in TE1 cells of si-DGCR5 group decreased significantly (P<0.01). Conclusion: lncRNA DGCR5 is highly expressed in ESCC cells, and promotes the proliferation, invasion and migration of TE1 cells possibly by up-regulating EGFR expression.

Objective: To investigate the effect and mechanism of bridging intergrator-1 (BIN1) on expression of programmed death-ligand 1 (PD-L1) in non-small cell lung cancer (NSCLC) A549 cells. Methods: Quantitative real-time polymerase chain reaction (qRTPCR) and Western blotting were used to detect the mRNA and protein expression of BIN1 and PD-L1 in A549 cells and normal human embryo lung fibroblast 2BS cells, respectively. Eukaryotic expression plasmid CMV-MCS-GFP-SV40-Neomycin-BIN1 containing human full length BIN1 gene sequence was transfected into A549 cells via cationic liposomes by using gene transfection technology (as BIN1+group); c-MYC-siRNAwas used to knockdown the expression of c-MYC inA549 cells through RNAinterference technique (as cMYC-siRNA group). The transfection efficiencies were verified by qRT-PCR and Western blotting, the effects of BIN1 over-expression and c-MYC knock-down on the expression of c-MYC and PD-L1 in A549 cells were detected as well. Results: Comparing with 2BS cells, the expression of BIN1 was down-regulated in A549 cells at both mRNA and protein levels, while the expression of PD-L1 was up-regulated (all P<0.05). The expression of BIN1 was increased at both mRNA and protein level in BIN1+ group, while the expression of PD-L1 was decreased significantly after B1N1 transfection (all P<0.05). After transfection of c-MYC-siRNA into A549 cells, the expression of c-MYC and PD-L1 in c-MYC-siRNAgroup was down-regulated significantly (all P<0.01). Conclusion: Over-expression of BIN1 could reduce the expression of PD-L1 by inactivating the c-MYC pathway, thereby inhibiting the immune escape ofA549 cells.

[摘 要] 目的：探讨PD-1抗体联合化疗对比抗血管生成药物联合化疗在晚期驱动基因阴性肺腺癌一线治疗中的疗效和安全性。方法：收集2018年3月至2021年8月河北医科大学第四医院收治的141例不可手术切除的ⅢB/ⅢC和Ⅳ期驱动基因阴性肺腺癌患者，回顾性分析PD-1抗体联合化疗对比抗血管生成药物联合化疗在一线治疗中的疗效与安全性。主要研究终点为无进展生存期（PFS），次要终点为客观缓解率（ORR）、疾病控制率（DCR）和不良反应。结果：141例患者均纳入生存分析，中位随访时间为13.0个月（95% CI：12.0～14.0）。PD-1抗体联合化疗组（A组）和抗血管生成药物联合化疗组（B组）的ORR分别为33.33%和27.38%，DCR分别为98.25%和89.29%，差异均无统计学意义。A组和B组的中位PFS分别为8.4个月（95% CI: 7.3～9.9）和6.9个月（95% CI: 6.1～7.7），差异无统计学意义。亚组分析结果显示，ⅢB/ⅢC期、肝或脑转移患者中，A组中位PFS较B组均延长（均P<0.01）。A组和B组不良反应发生率分别为26.32%和14.29%，多数为1~2级。结论：PD-1抗体联合化疗对比抗血管生成药物联合化疗一线治疗晚期驱动基因阴性肺腺癌疗效相当，不良反应可耐受，可成为晚期驱动基因阴性肺腺癌标准一线治疗。