The variable domain of heavy-chain antibody (VHH) has been developed widely in drug therapy, diagnosis, and research. Escherichia coli is the most popular expression system for VHH production, whereas low bioactivity occurs sometimes. Mammalian cells are one of the most ideal hosts for VHH expression at present. To improve the yield of VHH in Expi293F cells, we optimized the signal peptide (SP) and codons of VHH. Firstly, the fusion protein VHH1-Fc was used to screen SPs. The SP IFN-α2 showed the highest secretion as quantified by enzyme-linked immunosorbent assay (ELISA). Subsequently, codon optimization by improving GC3 and GC content doubled the yield of VHH1 and kept its binding activity to Senecavirus A (SVA). Finally, the mean yields of other 5 VHHs that fused with SP IFN-α2 and codon-optimized were over 191.6 mg/L, and these VHHs had high recovery and high purity in the culture supernatant. This study confirms that SP IFN-α2 and codon optimization could produce VHHs in Expi293F cells efficiently, which provides a reference for the large-scale production of VHHs.
Codon/genetics* ; Protein Sorting Signals/genetics* ; Escherichia coli/metabolism* ; Humans ; Recombinant Fusion Proteins/biosynthesis* ; Interferon-alpha/metabolism* ; Immunoglobulin Heavy Chains/immunology* ; Cell Line ; Immunoglobulin Variable Region/immunology*

Objective:To evaluate the necessity, safety and efficacy of endovascular treatment for cerebral infarction caused by middle cerebral artery (MCA) stenosis with hypoperfusion in the blood supply area of the lenticulostriate artery.Methods:The clinical and surgical data of patients with MCA atherosclerotic disease who underwent endovascular treatment in the First Affiliated Hospital of Zhengzhou University from January 2014 to October 2021 were retrospectively analyzed. A total of 6 patients with cerebral infarction caused by MCA stenosis with hypoperfusion in the blood supply area of the lenticulostriate artery were selected. The preoperative and postoperative clinical imaging characteristics, perioperative complications and follow-up of these 6 patients were summarized and evaluated.Results:After the endovascular treatment, the imaging of the lenticulostriate artery in all the 6 patients was clearer than that before the operation, and the number of main trunks of the lenticulostriate artery shown by imaging in 2 patients was more than that before operation. The computer tomography perfusion of 6 patients after the endovascular treatment showed that perfusion in the supply area of the lenticulostriate artery was significantly improved compared with pre-operation. No stroke, transient ischemic attack (TIA) and death occurred during the perioperative period. The time of clinical follow-up was 360 (322, 495) days, and there were no stroke, TIA or death occurring in the corresponding artery. All the 6 patients underwent imaging follow-up, of which 3 patients underwent digital subtraction angiography and 3 underwent CT angiography. The lumen of the target vessels showed patency in all patients.Conclusions:With rigorous imaging evaluation, endovascular treatment may be safe and effective for cerebral infarction caused by MCA stenosis with hypoperfusion in the blood supply area of the lenticulostriate artery.

Injection fear is widespread in the population, which can cause patients to tolerate or avoid injection, reduce treatment compliance, and increase the burden of healthcare. Choosing appropriate injection fear assessment tools in clinical practice is helpful to understand the degree, psychological characteristics and influencing factors of individual injection fear. In this paper, the contents, characteristics and application methods of fear of injection assessment tools at home and abroad are reviewed, in order to provide reference for the application and development of fear of injection assessment tools for medical staff.

Objective    To investigate the effects of telmisartan on the proliferation, migration and apoptosis of non-small cell lung cancer A549 and the mechanism of regulating Wnt signaling pathway. Methods    Non-small cell lung cancer cell line A549 was cultured in vitro. Cell counting kit-8 (CCK-8) assay was used to detect the effect of telmisartan at different concentrations on the proliferative activity of A549 cells. The survival fraction of A549 treated with different concentrations of telmisartan was determined by colony-formation assay. The effect of telmisartan at different concentrations on the migration ability of A549 cells was examined in the wounding healing assay. Hoechst staining was used to detect the effects of telmisartan at different concentrations on the apoptosis of A549. Western bloting was used to detect the expressions of β-actin, proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, Wnt-3a, Beta-catenin (β-catenin), serine protein kinase 3β (p-GSK-3β), glycogen synthase kinase-3β (GSK-3β) and c-myc. Results    Different concentrations of telmisartan treatment inhibited the proliferation activity, colony-formation rate and migration of A549 cells, and reduced the expression of PCNA in a concentration-dependent manner. Telmisartan treatment promoted the apoptosis of A549 cells, significantly increased the expression of pro-apoptotic protein Bax and decreased the expression of anti-apoptotic protein Bcl-2. The expression levels of Wnt-3a, β-catenin, p-GSK-3β, and c-myc in A549 cells increased after treatment with telmisartan, while the expression levels of GSK-3β decreased. Conclusion    Telmisartan may play a role in the proliferation, migration and apoptosis of non-small cell lung cancer A549 cells, and  inhibiting the Wnt/β-catenin signaling pathway may be one of the mechanisms.

Purpose: Anterior gradient 3 (AGR3) belongs to human anterior gradient (AGR) family. The function of AGR3 on cancer remains unknown. This research aimed to investigate if AGR3 had prognostic values in invasive ductal carcinoma (IDC) of breast cancer and could promote tumor progression. Materials and Methods: AGR3 expression was detected in breast benign lesions, ductal carcinoma in situ and IDC by immunohistochemistry analysis. AGR3’s correlations with clinicopathological features and prognosis of IDC patients were analyzed. By cell function experiments, collagen gel droplet-embedded culture drug sensitivity test and cytotoxic analysis, AGR3’s impacts on proliferation, invasion ability, and chemotherapeutic drug sensitivity of breast cancer cells were also detected. Results: AGR3 was up-regulated in luminal subtype of histological grade I-II of IDC patients and positively correlated with high risks of recurrence and distant metastasis. AGR3 high expression could lead to bone or liver metastasis and predict poor prognosis of luminal B. In cell lines, AGR3 could promote proliferation and invasion ability of breast cancer cells which were consistent with clinical analysis. Besides, AGR3 could indicate poor prognosis of breast cancer patients treated with taxane but a favorable prognosis with 5-fluoropyrimidines. And breast cancer cells with AGR3 high expression were resistant to taxane but sensitive to 5-fluoropyrimidines. Conclusion AGR3 might be a potential prognostic indicator in luminal B subtype of IDC patients of histological grade I-II. And patients with AGR3 high expression should be treated with chemotherapy regimens consisting of 5-fluoropyrimidines but no taxane.

Objective To reveal the virulence genes and the polymorphisms of chromosomal 16S rRNA gene of Yersinia enterocolitic strains isolated from different districts in Jiangsu Province,2015. Methods Five virulence genes(ail,virF,yadA,ystA and ystB)of Yersinia enterocolitic strains isolated from different districts in Jiangsu Province were detected by using polymerase chain reaction(PCR),and phylogenetic analysis of chromosomal 16S rRNA gene was performed by amplification and sequencing. Results In this study,73 Yersinia enterocolitic strains were collected in Jiangsu Province in 2015.Among them,56(76.7%)strains carried virulence genes,and ail-virF-yadA -ystA -ystB+were the dominate types in diarrhea patients and other hosts.All strains can be clustering into 4 groups according to the phylogenetic analysis of chromosomal 16S rRNA gene.Conclusions The non-pathogenic Yersinia enterocolitic(ystB+)is the dominant strain in Jiangsu province,and the pathogenic strains are also found in this region.The result of phylogenetic analysis of chromosomal 16S rRNA gene and the profiles of virulence genes are highly consistent.

The 13th World Congress of the International Hepato-Pancreato-Biliary Association was held from 4th to 7th September 2018 in Geneva,Switzerland.Thousands of wellknown specialists and scholars from 96 countries around the world were invited for the great event.The congress aimed to explore the latest achievements of diagnosis and treatment of hepatobiliary and pancreatic diseases from both the clinical and basic perspective.In this article,authors reviewed and analyzed the up-to-date research information and combined clinical researches of the author team,in order to share the experience and achievements in the field of hepato-pancreato-biliary surgery with colleagues and provide new information and inference for optimization of diagnosis and treatment in this field.

Objective To discuss the application effect of zero OT-U type bionics urine bag in neurology critically ill patients to reduce the indwelling catheterization complications. Methods A total of 50 cases of critically ill patients with indwelling catheter were chosen from July 2014 to May 2015. They were randomly divided into the experimental group and the control group with 25 cases in each group according to the time of admission. The control group used common urine bags and the experimental group adopted bionics urine bag. The excessive urine, urinary tract infection, incontinence associated dermatitis, the time with indwelling urinary catheter,replacing the urine tube to urinate in two groups were compared. Results The rate of excessive urine, urinary tract infection, incontinence associated dermatitis and the time with indwelling urinary catheter in the experimental group were 12%(3/25), 4%(1/25), 32%(8/25) and (10.120 ±3.295) days, while the results were 84%(21/25), 48%(12/25), 80%(20/25) and (28.040±9.480) days in the control group, the differences were different (χ2=27.931,7.741, 7.018, t=8.927, P<0.01). The cases of voluntary micturition, induced micturition and secondary catheterization were 17, 5 and 3, and the results in the control group were 3, 12 and 10, the differences were were different ( χ2=16.452, P < 0.01). Conclusions The zero OT-U type bionics urine bag has a bigger superiority in reducing the related complications in neurology critically ill patients with indwelling catheter. At the same time , the zero OT-U type bionics urine bag reduced the patient's pain, improved the family satisfaction and reduced the pressure on care and treatment.

Objective To investigate the influencing factors for trauma-induced tibia1 infection in underground coal mine.Methods A retrospective analysis was performed for the clinical data of 1 090 patients with tibial fracture complicated by bone infection who were injured in underground coal mine and admitted to our hospital from January 1995 to August 2015,including the type of trauma,injured parts,severity,and treatment outcome.The association between risk factors and infection was analyzed.Results Among the 1 090 patients,357 had the clinical manifestations of acute and chronic bone infection,219 had red and swollen legs with heat pain,and 138 experienced skin necrosis,rupture,and discharge of pus.The incidence rates of tibial infection from 1995 to 2001,from 2002 to 2008,and from 2009 to 2015 were 31％,26.9％,and 20.2％,respectively.The incidence rate of bone infection in the proximal segment of the tibia was significantly higher than that in the middle and distal segments (42.1％ vs 18.9％/27.1％,P＜0.01).As for patients with different types of trauma (Gustilo typing),the patients with type Ⅲ fracture had a significantly higher incidence rate of bone infection than those with type Ⅰ/Ⅱ infection (52.8％ vs 21.8％/24.6％,P＜0.01).The incidence rates of bone infection after bone traction,internal fixation with steel plates,fixation with external fixator,and fixation with intramedullary nail were 20.7％,43.5％,21.4％,and 26.1％,respectively,suggesting that internal fixation with steel plates had a significantly higher incidence rate of bone infection than other fixation methods(P＜0.01).The muhivariate logistic regression analysis showed that the position of tibial fracture and type of fracture were independent risk factors for bone infection.Conclusion There is a high incidence rate of trauma-induced tibial infection in workers in underground coal mine.The position of tibial fracture and type of fracture are independent risk factors for bone infection.Vacuum sealing drainage and Ilizarov technique can achieve a satisfactory therapeutic effect.