Objective To express recombinant human interleukin-29-Fc(rhIL-29-Fc) fusion protein in human embryonic kidney 293-F(HEK293F) cells and analyze its anti-tumor activity in vitro.Methods The recombinant expression plasmid UCOE-IL-29-Fc was constructed and transiently transfected into HEK-293F cells.After expression and purification,rhIL-29-Fc fusion protein was obtained and identified by SDS-PAGE and Western blot;Female Japanese white rabbits were immunized with rhIL-29 and rhIL-29-Fc protein subcutaneously in the left ear respectively,2 rabbits in each group,0.5 mg per rabbit.Blood samples were collected from the vein of right ear,and the serum was separated.The half-life was measured by ELISA and the anti-proliferation effect of rhIL-29-Fc protein on human colon cancer HT-29,human colon cancer HCT-116,human Burkkit lymphoma Daudi,human non-small cell lung cancer NCI-H1975,human small cell lung cancer NCI-H209,human esophageal cancer EC109 and human pancreatic cancer PANC-1 cells in vitro was detected by CCK-8 assay,and the inhibitory concentration 50(IC_(50)) was calculated.Results The recombinant expression plasmid UCOE-IL-29-Fc was constructed correctly as identified by double digestion and sequencing.After transient transfection into HEK-293 cells for 6 d,the culture supernatant was harvested.The relative molecular mass of the purified rhIL-29-Fc fusion protein was consistent with the expectation.The protein showed a specific binding reaction with mouse anti-human IL-29 monoclonal antibody with a concentration of 1.5 mg/ml and a purity of 93%.RhIL-29-Fc protein had a half-life of 25 h and showed different inhibitory effects on the proliferation of 7 kinds of tumor cells,and the IC_(50) on different cells was also different.Conclusion The rhIL-29-Fc fusion protein was successfully expressed in HEK-293F cells,and the half-life of the fusion protein was 20 h longer than that of rhIL-29.According to the different anti-tumor proliferation activity in vitro and IC_(50) results on 7 kinds of tumor cells,it was found that the anti-tumor activity of rhIL-29-Fc fusion protein was higher than that of rhIL-29.This study laid a foundation of the development of IL-29 protein in the treatment of tumors.

Objective To express recombinant human interferon λ1(rhIFNλ1)by transient transfection in HEK-293F cells and identify it. Methods Two signal peptides[T cell receptor(TCR)and nature signal peptide(NSP)],three vectors[pcDNA3.4,ubiquitous chromatin opening element(UCOE)and PFR]and the target gene rhIFNλ1 were used to construct recombinant plasmids of six signal peptide-vector combinations. Using HEK-293F as host cells,the recombinant plasmids were transfected transiently to express rhIFNλ1 on a shake flask scale. The recombinant plasmid UCOE-Q46-λ with low glycosylation rhIFNλ1 was constructed by using NSP and vector UCOE,and transfected transiently into HEK-293F cells. The expressed product was purified by cation exchange chromatography(HiTrap SP FF),blue gel chromatography(HiTrap Blue HP)and gel filtration chromatography(Sephacryl S-100 HR),which was then analyzed by Western blot and reversed-phase HPLC,and determined for its molecular mass and N-terminal amino acid sequence by mass spectrometry. Results Six recombinant plasmids were constructed correctly as identified by double enzyme digestion and sequencing. With the extension of transfection time,the expression levels of the six expressed products increased gradually,and reached the highest level of10 ～ 20 mg/L at the 6th day of transfection. Double enzyme digestion and sequencing identification proved that the recombinant plasmid UCOE-Q46-λ with low glycosylation rhIFNλ1 was constructed correctly. The recombinant plasmid UCOE-Q46-λwas transfected into HEK-293F cells for 6 d. The purified product of cell culture supernatant showed a relative molecular mass of about 27 800 and a purity of 97. 372%,which showed specific binding to mouse anti-human IL-29/IFNλ1 monoclonal antibody;Two peaks were detected by reversed-phase HPLC,and the peak was showed at 15. 6 min and 20. 0 min respectively;The mass spectrometry molecular mass was about 24 000;The N-terminal five amino acids were G-P-V-P-T.Conclusion The rhIFNλ1 expressed by HEK-293F cells has high purity,which lays a foundation of further study of the protein.

Objective To describe our follow-up results of interventional management in 20 patients with subclavian artery stenosis. Methods This study involved in 20 symptomatic patients with an average age of 63.4 years,a mean stenose severity of 86.2% and a mean systolic arterial pressure difference of 116 mm?Hg between both upper limbs. Percutaneous transluminal angioplasty (PTA) in the first 8 patients and stenting in the last 12 patients were performed via a right femoral or radial artery. Results Both PTA and stenting were successfully archived in all patients with normalization of radial pulses and a mean systolic arterial pressure difference of 24 mm?Hg between both upper limbs. No major complications occurred. In the follow-up for an average of 18.5 months,19 patients were free of adverse events except one with symptom recurrence. Conclusion As effective,simple and safe procedures,PTA and stenting can be used as a first-line treatment modality for the symptomatic patients with subclavian artery stenosis.

cancer,and prostatic cancer has been reported.

This paper analyzed medical dispute litigation from the view of game theory.The adoption of mediation strategy for both hospital and patient was the Pareto optimal of this model.The adoption of court decision for both hospital and patient was the Nash equilibrium and its overall benefit was the Pareto suboptimal of this model.The communication of patient's lawyer can prompt the reconciliation between hospital and patient.On the one hand,it let the hospital realize the responsibilities they should bear,on the other hand let patients be willing to make concessions on the basis of the original claims,and finally made the two sides reach a reconciliation agreement,thus to make conflict between hospital and patient end in litigation and the overall benefits maximize.

Objective To observe the frequency of interleukin-6(IL-6) gene -572C/G polymorphism and its relationship with myocardial infarction(MI) and serum lipids in Chinese Hans. Methods IL-6/gene-572C/G polymorphism was genotyped in 232 MI patients and 260 healthy adults by PCR-RFLP method. Results There were IL-6/gene-572 CC、CG and GG genotype. -572GG genotype and G allele were more frequent in patients than those in controls(P

AIM:This paper described a HPLC method.Paeoniflorin and puerarin in Rg soft-exact(Radix Puerariae lobatae,Redix Paeoniae alba,etc.) were simultaneously determined.METHODS:A simplified HPLC procedure was developed.A 4.6 mm?250 mm Alltima C_(18) column with a mobile phase consisting of NaH_2PO_4(20 Mm,pH 2.4)-Acetonitrile:Tetrahydrofuran(900:120:5) was used.The flow rate was 1.00 mL/min.It was detected at 320 nm.RESULTS:Paeoniflorin and puerarin were well separated.The average recoveries of paeoniflorin and puerarin were 98.10% and 101.2% respectively.CONCLUSION:The result shows that this method is convenient,rapid and accurate.

Objective To investigate the expression of minichromosome maintenance protein 7 (MCM7) in sinonasal squamous cell carcinoma (SNSCC),and evaluate its relationship with tumor differentiation and prognosis of patients.Methods Using immunohistochemistry,MCM7 expression in SNSCC and nasal polyps with chronic sinusitis (NPCRS) were studied,and relationships between markers and clinicapathological features were analyzed.Results In NPCRS,MCM7 positive cells were mainly distributed in the epithelial basal layer and the expression rate was low,whereas in SNSCC,MCM7 positive cells were diffuse and the expression rate was high.MCM7 expression was significantly higher in SNSCC than in NPCRS (P ＜ 0.001) and related to tumor differentiation (P =0.001),increasing gradually with decreasing degree of differentiation.The overall 3-and 5-year survival rates of patients with SNSCC were 61.3％ and 46％,respectively.The 3-year survival rates for patients with stage Ⅰ and Ⅳ were 90％ and 25.6％,respectively,and the 5-year survival rates were 70％ and 17.1％,respectively;the difference was statistically significant (P ＜ 0.001).The 3-and 5-year survival rates of MCM7-negative patients were 36.0％ and 18.0％,respectively,and those of MCM7-positive patients were 59.9％ and 34.2％,respectively;the difference was not statistically significant (P =0.297).Conclusion In SNSCC,MCM7 expression is significantly increased,inversely associated with the degree of tumor differeutiation,and unrelated to the survival rates of patients.

Objective: To investigate the development of osteoporosis (OS) and its influencing factors among physical examination populations aged over 35 years in urban areas of Hangzhou City, so as to provide insights into OP prevention. Methods: Subjects aged over 35 years that underwent physical examinations in Wenhui Street Community Health Service Center of Gongshu District, Hanghzhou City during the period between April and July, 2022 were enrolled. Subjects' demographic characteristics and physical examination data were collected, and the bone mineral density and serum vitamin D levels were measured. The factors affecting the development of OP were identified using a logistic regression model. Results: A total of 1 561 subjects were recruited, including 616 males (39.46%) with a median (interquartile range) age of 62 (18) years, and 945 females (60.54%) with a median (interquartile range) age of 62 (19) years. The prevalence of OP was 43.69% among the study subjects. Multivariable logistic regression analysis identified women (OR=1.367, 95%CI: 1.105-2.979) and increased age (OR=1.057, 95%CI: 1.044-1.069) as risk factors for OP and elevated serum 25-hydroxy-vitamin D levels as a protective factor for OP (OR=0.969, 95%CI: 0.954-0.985). Conclusions The prevalence of OP was 43.69% among physical examination populations aged over 35 years in urban areas of Hangzhou City, and gender, age and serum 25-hydroxy-vitamin D were factors affecting the development of OP.

Objective:To explore the effect of spinal interleutkin-33 (IL-33) on radicular pain in rats modeling non-compressive lumbar disc herniation.Methods:A total of 80 male Sprague-Dawley rats were randomly divided into a sham operation group, a model group, a lentivirus negative control group, a low-dose IL-33 recombinant lentivirus group and a high-dose IL-33 group, each of 16. Non-compressive lumbar disc herniation was successfully induced in all except the rats in the sham operation group. Two days later, the model group was injected intrathecally with 10μl of enhanced infection solution. The lentivirus control group received 10μl of negative lentivirus, the low-dose IL-33 recombinant lentivirus group received 5μl of IL-33 recombinant lentivirus and the high-dose IL-33 recombinant lentivirus group received 10μl of IL-33 recombinant lentivirus. The 50% paw withdrawal threshold (50% PWT) was measured one day before the modeling and on the 1 st, 3 rd, 5 th, 7 th, 9 th, 11 th, 13 th, 15 th, 17 th, 19 th, and 21 st day afterward. On the 12 th day the expressions of IL-33 protein and mRNA were evaluated. Results:The average expression of IL-33 protein and mRNA in the model and the lentivirus negative control group increased significantly after the modelling compared with the sham group, while expression in the low- and high-dose IL-33 recombinant lentivirus groups was significantly lower than in the lentivirus negative control group. Compared with one day before the modelling, average 50% PWTs on the affected side decreased significantly in all of the modelling groups. From the 9 th to the 21 st day significantly increased 50% PWTs were observed on the affected side in the low-dose and high-dose IL-33 recombinant lentivirus groups compared with the other two modelling groups. Immunostaining showed significant increase in the expression of IL-33 in the dorsal horn of the spinal cord in the model group, compared with the sham operation group. Significant decrease in the average expression of IL-33 in the spinal dorsal horns was observed in the low-dose and high-dose IL-33 recombinant lentivirus groups. Conclusions:Intervertebral disk herniation may increase the expression of IL-33 in the spinal cord, and may cause radicular pain.