Development and verification of a rapid detection method for adalimumab biological activity based on U937-NF-κB-Luc cell line / 中国生物制品学杂志

@#Objective To develop and verify a rapid detection method for the biological activity of adalimumab based on U937-NF-κB-Luc cell line. Methods Using U937-NF-κB-Luc cell line as the detection cells,a method for detecting the biological activity of adalimumab was developed based on luciferase luminescence principle. The method was optimized for the concentration of tumor necrosis factor-α(TNF-α)(160 ng/mL as initial concentration,2 times serial dilution,10dilutions),the initial concentration of antibody(2 000 ng/mL,2 times serial dilution,20 dilutions),the dilution multiple of antibody(1. 5,2,3,4 times),the inoculation amount(8 × 103,2 × 104,4 × 104,6 × 104cells/well)and the incubation time(0. 5,1,2,3 h),and verified for the specificity,accuracy,precision and linear range. The relative potency of five batches of adalimumab was detected by using the optimized method and TNF-α neutralization activity method based on L929cells respectively. Results The dose-response curve of adalimumab international standard showed a typical S-type,and the data complied with the four-parameter equation y =(A-D)/[1 +(x/C)B]+ D,R2> 0. 99. The optimum concentration of TNF-α was 5 ng/mL,the initial concentration of antibody was 800 ng/mL,the dilution ratio for adalimumab was 1∶2,the inoculation amount was 2 × 104cells/well,and the induction time was 2 h. Three therapeutic monoclonal antibodies of TNF-α target,such as adalimumab,obtained good dose-response curves,while therapeutic monoclonal antibodies of other non-TNF-α targets did not show this curve. The linear regression equation of the logarithmic value of theoretical potency and the logarithmic value of the corresponding measured potency had a slope of 1. 037,and the relative bias was within the range of ± 12%. The geometric coefficient of variation(GCV)of the relative titer measured value of each sample was less than20%. The theoretical potency ranged from 64% to 156%,showing a good linear relationship with the measured values,and the fitting linear regression equation was y = 1. 037 4 x-0. 023 7,R2= 0. 998 4. There was no significant difference in the relative potency measured results of five batches of adalimumab by the two methods(t = 1. 198,P = 0. 265 1). Conclusion The developed detection method for adalimumab biological activity based on U937-NF-κB-Luc cell line has good specificity,accuracy and precision with short time consumption(3 h),which can be used as a rapid evaluation method for the biological activity of adalimumab.

Comparison with efficacy and safety of the treatment with different operation approach under nasal endoscope in benign maxillary sinus lesions / 中国内镜杂志

Objective To explore the validity and security of treatment of different operation approach under nasal endoscope in patients with benign maxillary sinus lesions.Methods A retrospective analysis was performed on 80 patients with benign maxillary sinus lesions diagnosed and treated from January 2020 to January 2022,and they were divided into group A(n = 40)and group B(n = 40)according to the operative method.Group A was treated with middle nasal meatus combined with Caldwell-Luc operation under nasal endoscope,and group B was treated with middle nasal meatus antrostomy first and anterior lacrimal recess approach later under nasal endoscope.The clinical efficacy,operation index,visual analogue scale(VAS),Lund-Kennedy endoscopic score,adverse reactions and recurrence rate of both groups were compared before and after operation.Results The total effective rate was sensibly higher in group B than that in group A,the intraoperative blood loss was sensibly less in group B than that in group A,the operation and hospitalization time were sensibly shorter in group B than those in group A,6 months after operation,the VAS and Lund-Kennedy scores were sensibly decreased in comparison with those before operation,and group B was sensibly decreased in comparison with those in group A,the total incidence rate of adverse reactions and recurrence rate were sensibly lower in group B than those in group A,the differences were statistically significant(P＜0.05).Conclusion The surgical treatment of patients with benign maxillary sinus lesions through the middle nasal meatus and the anterior lacrimal recess approach under nasal endoscope is effective and can relieve the pain of patients with high safety.It is worthy of clinical application.

Establishment of mouse models of lymphoma with dual-labeled EBV-immortalized lymphoblastoid cell lines by intravenous versus subcutaneous injection / 中国肿瘤临床

Objective:To establish a green fluorescent protein(GFP)and firefly luciferase(Luc)double-labeled Epstein-Barr virus(EBV)infec-ted B lymphoblastoid cell lines(B-LCL)and apply them to mouse models,then compare the advantages and disadvantages of models inocu-lated by intravenous(IV)or subcutaneous(SC).Methods:B lymphoblastoid cell lines double-tagged with GFP/Luc(B-LCL-GL)were con-structed through lentivirus transduction,puromycin intervention.Subcutaneous xenograft and hematogenous metastasis models were re-spectively established by subcutaneous or intravenous injection of B-LCL-GL cells at three concentrations in(NOD)/Prkdcscid/IL-2Rγnull(NPG)mice for in vivo bioluminescence imaging.Results:In the B-LCL-GL group,the ratio of the GFP-positive cell population was 92.5%,and the average luminescence intensity was as high as 4.80E+08 Photons/s,which was considerably higher than that of untreated B-LCLs.In the hematogenous metastasis models,tumor bioluminescence was initially located in the peritoneal area and then spread throughout the en-tire body between 7 and 28 days.In the subcutaneous xenograft models,strong central and weak peripheral tumor-related biolumines-cence signal was detected on day 7 in the three groups,which then spread throughout the body on day 28 in the high-dose group.Taken to-gether,there was no significant difference in tumor progression between the two routes of administration when using the same dose of B-LCL-GL cells.However,the survival analysis indicated that the IV injection group,in which all the mice ultimately died,had a shorter time frame for testing than that of the SC injection group,in which the mice survived until day 100 in the low-dose and medium-dose groups,thus allowing for long-term testing.Conclusions:GFP and Luc dual-positive B-LCLs were successfully established to generate hematogenous metastasis and subcutaneous xenograft models,which allow the monitoring of the location and size of lymphomas in vivo.It provide plat-form for the study of tumor characteristics and selecting anti-tumor drugs.

Luc’s abscess: The zygomatic route of infection from cholesteatoma

@#Luc’s abscess is an uncommon complication of otitis media wherein a subperiosteal abscess develops into the temporalis muscle and follows the route of a pneumatized zygoma.1 In uncomplicated cases, surgical drainage and antibiotics are adequate management with mastoidectomy reserved for severe or complicated cases. We report a case of complicated Luc’s abscess presenting with many complications that required multiple surgical interventions.

Antidiabetic Activity of Aqueous and Ethanolic Extracts of Stem Barks from Trichilia emetica (Meliaceae) in Alloxan-Induced Diabeticalbinos Rats

Aims: This study was designed to evaluate the antidiabetic effects of aqueous and ethanolic extracts of Trichilia emetica(TE) stem bark in albinos rats.Methodology:24 rats of comparable weight were divided into 2 lots, including a control lot of 3 rats and a test lot of 21 rats. Diabetes was induced by injecting Alloxan® intraperitoneally (125 mg/kg bw) for 7 days. After the onset of diabetes, the animals were divided into several groups and given the extracts of Trichilia emeticaand Diastabol®orally for 6 days according to the following protocol: Normal Control (NC) received normal saline, group diabetic control (DC) it consists of untreated diabetic, group DD10 and DD20 was comprised of diabetic animals treated with Diastabol® at a doses of 10 and 20 mg/kg bw respectively , group DTEE100 and DTEE200 it consists of diabetic animals treated with ethanolic extract at a doses of 100 and 200 mg/kg bw respectively,group DTEA100 and DDTEA200 was comprised of diabetic animals treated with aqueous extract at a same dose (100 and 200 mg/kg bw). At the end of experimentation, some blood was collected for the determination of some biochemical parameters such as insulin, blood glucose, AST, ALT, urea and creatinine. Systolic blood pressure (SBP), diastolic blood pressure (DBP), cardiac frequency (CF) were also recorded. Results:The results showed a significant (P<0.05) decrease in insulin levelsin diabetic rats compared with NC. In addition, diabetes caused a significant increase (P<0.05) in blood glucose, urea, creatinine levels,transaminase activity, and in blood pressure numbers (DBP, SBP and CF) still in comparison with NC. However, the treatment of sick animals with the extracts and the Diastabol at the doses listed above significantly (P <0.05)increased insulin levels and reduced the sames biochemical parameters levels as well as the blood pressure numberscompared with DC.Conclusion:The data obtained showed that this part of the plant would have antidiabetic effects with ethanolic extract at the dose of 200 mg/kg bw had the highest pronounced effect and could be used as a good alternative for diabetes management, thus justifying its use in traditional medicine

Comparison Between Endoscopic Prelacrimal Medial Maxillectomy and Caldwell-Luc Approach for Benign Maxillary Sinus Tumors

OBJECTIVES: Endoscopic prelacrimal medial maxillectomy (EPMM) was previously reported to treat maxillary inverted papilloma. This study aimed to compare prelacrimal recess approach with the conventional Caldwell-Luc approach (CLA) to remove benign maxillary sinus tumors and to evaluate the usefulness of this approach based on our experience. METHODS: Ten patients who underwent EPMM at our hospital from January 2013 to December 2017 were reviewed. We also reviewed 30 patients who underwent benign maxillary sinus tumor resection via CLA during the same period. From medical records, postoperative pathological results, complications due to surgery, and recurrence rate were evaluated. RESULTS: There were eight inverted papilloma, one ameloblastoma, and one ossifying fibroma in the EPMM group. In the CLA group, all 30 cases were inverted papilloma. There were no cases of failure at gross total removal during surgery, and no recurrences were observed during follow-up in either groups. Mean follow-up period was 13.0 months in CLA group and 10.8 months in EPMM group. Regarding postoperative complications, 11 patients of the CLA group (37%) and three patients of the EPMM group (30%) had numbness around the cheek and upper lip area after surgery (P=0.715). In the CLA group, there were eight patients who had numbness lasting more than 3 months after surgery, and two patients had numbness for more than 1 year. However, facial numbness disappeared within 3 months in all patients in the EPMM group, in which epiphora was not observed. CONCLUSION: EPMM is the effective surgical approach for resecting benign maxillary sinus tumor compared with CLA. Although facial numbness was reported in EPMM, the duration of numbness was shorter than CLA.

Treatment of Facial Neuralgia Developed after Inferior Meatal Antrostomy by Narrowing of the Inlet with Endoscopic Cartilage Graft

Inferior meatal antrostomy (IMA) is a widely performed surgical technique to treat postoperative maxillary mucocele. The method is safe and easy to perform, without major complications compared with other approaches. Facial pain after IMA is a rare clinical entity that can be challenging to diagnose and treat. The authors present an unusual case of acute facial neuralgia triggered by cold air that developed after IMA. The antrostomy was located at the anterior-most part of the inferior meatus, and the inlet size was relatively large compared with the size of the remaining sinus. Surgical narrowing of the antrostomy inlet using endoscopy dramatically reduced the symptoms, and symptom relief was maintained for up to one year after surgery.

Inhibitory effect of recombinant oncolytic adenovirus on luciferase-labeled and non-labeled human lung cancerA549 cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the inhibitory effect of recombinant oncolytic adenovirusAd-Apoptin-hTERTp-E1A(ATV) on luciferase-labeled human lung cancer cells (A549-luc) and human lung cancerA549 cells, and to compare the differences in the inhibitory effect on two cell lines. Methods:ATV was used to infectA549-luc cells andA549 cells respectively. WST-1 and crystal violet staining were used to determine the difference in the inhibitory effect of ATV. Hoechst and Annexin V-FITC/PI staining were used to verify the inhibition mode ofATV. Results: WST-1 and crystal violet staining showed thatATV had significant inhibitory effect on bothA549-luc and A549 cells ( P <0.05). ATV showed significant inhibitory effect on both cells at 24, 48 and 72 h ( P <0.05 or P <0.01), and reached the peak at 72 h; ATV at concentrations of 1, 10 and 100 MOI all showed inhibitory effect on both cells, and reached the peak at 100 MOI. Hoechst staining showed that A549-luc cells and A549 cells infected with ATV showed typical nuclear fragmentation and marginal set. The results of Annexin V-FITC/PI Flow cytometry showed that ATV infection resulted in apoptosis of A549-luc and A549 cells, which was in a time-dependent manner and reached the peak at 72 h( P <0.05 or P <0.01). Conclusion: Insertion of luciferase didn’t significantly change the inhibitory effect and inhibitory mode ofATV onA549-luc cells.ATV exerted its in vitro inhibitory effect onA549-luc and A549 cells by inducing cell apoptosis.

Luc Montagnier: sus investigaciones, la relación con la homeopatía y su contrastación con los estudios de los físicos Poponin, Garaiev, Leikin y Popp / Luc Montagnier: his research, the relationship with homeopathy and its contrast with the studies of physicists Poponin, Garaiev, Leikin and Popp

En la búsqueda de una solución al problema del VIH, el profesor Luc Montagnier exploró la posibilidad de un co-factor utilizando las herramientas del doctor Jacques Benveniste y su equipo de trabajo; descubrió entonces que el líquido de los cultivos de Mycoplasma pirum contenía señales electromagnéticas (SEM) capaces de dar origen nuevamente a dichos microorganismos, en caso de que se depositaran linfocitos sanos en ese líquido. Posteriormente demostró que los virus y bacterias patógenas emiten SEM de 20 y 100 nm, respectivamente. Consiguió llevar a cabo una transducción del ADN enviando su señal electromagnética por internet a Italia y logrando su recuperación en un laboratorio de biología molecular. Emitió la hipótesis de la existencia de infecciones frías, responsables de, al parecer, muchas enfermedades no consideradas hasta la fecha como infecciosas, lo que representa un aporte valiosísimo a la Homeopatía pues concuerda con los señalamientos de Samuel Hahnemann sobre las enfermedades crónicas. Todo esto se sustenta en la existencia de nanoestructuras de agua, las cuales permiten que se conserven las SEM en su interior. Con base en estos descubrimientos experimentales, Montagnier ha señalado el peligro que representa vivir en un ambiente saturado de SEM y el aumento de personas electrosensibles. Así, ha establecido nuevos paradigmas y ha planteado si seremos capaces de adaptarnos a estos cambios tan rápidos. Estas propuestas se complementan y contrastan con los descubrimientos de físicos como Sergey Leikin, quien asegura que el ADN inicia no como una molécula, sino como una onda; Fritz-Albert Popp, descubridor del papel de los fotones en los procesos biológicos y quien asegura que estas subpartículas son esenciales para la salud y el funcionamiento del ADN, y Vladimir Poponin, quien descubrió el efecto fantasma del ADN, que no es otra cosa que su capacidad organizativa aún cuando las moléculas de ADN ya no están presentes.