Objective To screen the anti-CD22-specific nanobodies to provide a basis for immunotherapy agents. Methods The naive phage nanobody library was constructed and its diversity was analyzed. Three rounds of biotinylated streptavidin liquid phase screening were performed by using biotinylated CD22 antigen as the target, and the sequence of nanobodies against CD22 were identified by ELISA and gene sequencing. Results The capacity of the constructed naive phage nanobody library was 3.89×10⁹ CFU/mL, and the insertion of effective fragments was higher than 85%. Based on this library, seven anti-human CD22 nanobodies were screened, and the amino acid sequence comparison results showed that the overall similarity was 70.34%, and all of them were hydrophilic proteins. The results of protein-protein complex docking prediction showed that the mimetic proteins of the five nanobody sequences could be paired and linked to CD22, and the main forces were hydrophobic interaction and hydrogen bonding. Conclusion This study provided a basis for the study of chimeric antigen receptor T cells targeting CD22, successfully constructed the natural phage nanobody library and obtaining five anti-CD22-specific nanobodies.
Camelus/immunology* ; Single-Domain Antibodies/chemistry* ; Peptide Library ; Humans ; Animals ; Sialic Acid Binding Ig-like Lectin 2/genetics* ; Amino Acid Sequence ; Molecular Docking Simulation

Objective:To investigate the value of MRI combined with serum long non-coding RNA (lncRNA) KCNQ1OT1 and microRNA-204-5p (miR-204-5p) in predicting axillary lymph node metastasis in breast cancer patients.Methods:A total of 70 patients with breast cancer who were surgically diagnosed in the No.2 Hospital of Baoding, Hebei Province from January 2020 to December 2022 were selected as the study objects. Patients were divided into metastatic group ( n=31) and non-metastatic group ( n=39) according to the status of lymph node metastasis. The MRI features and the levels of serum lncRNA KCNQ1OT1 and miR-204-5p were compared between the two groups. Receiver operator characteristic (ROC) curve was used to evaluate the predictive efficacy of the three detections alone and in combination for axillary lymph node metastasis in breast cancer. Results:There were statistically significant differences in tumor maximum diameter ( χ2=4.28, P=0.039) and Ki-67 expression ( χ2=10.88, P=0.001) between the metastatic and non-metastatic groups. There were statistically significant differences between the breast cancer axillary lymph node metastatic group and the non-metastatic group in the mode of internal reinforcement type ( χ2=6.60, P=0.037), peripheral vessel diameter ( t=4.33, P<0.001), number of peripheral vessel roots ( t=4.38, P<0.001), apparent diffusion coefficient ( t=3.59, P=0.001), and MRI lymph node status ( χ2=29.70, P<0.001). The level of serum lncRNA KCNQ1OT1 in metastatic group was higher than that in non-metastatic group (1.41±0.32 vs. 0.99±0.18, t=6.94, P<0.001), and the level of miR-204-5p was lower than that in non-metastatic group (0.72±0.17 vs. 1.03±0.21, t=6.66, P<0.001). Bioinformatics analysis showed that miR-204-5p may have a targeting relationship with lncRNA KCNQ1OT1. ROC curve analysis showed the area under the curve of MRI features, serum levels of lncRNA KCNQ1OT1, miR-204-5p and their combined detection to predict lymph node metastasis of breast cancer patients were 0.823, 0.858, 0.843, 0.946, respectively. The predictive efficacy of the combined detection was better than that of MRI features and serum lncRNA KCNQ1OT1 and miR-204-5p levels alone ( Z=3.29, P=0.001; Z=2.07, P=0.038; Z=2.23, P=0.026) . Conclusions:Serum lncRNA KCNQ1OT1 level is up-regulated and the level of miR-204-5p is down-regulated in patients with axillary lymph node metastasis of breast cancer. And the combination of MRI features with serum lncRNA KCNQ1OT1 and miR-204-5p has high predictive efficacy in predicting lymph node metastasis in breast cancer patients.

Distal transradial access(dTRA)was first applied in coronary heart disease intervention by Kiemeneij in 2017,dTRA has become an important technological advancement of coronary interventional therapy.This approach,performed by puncturing the distal radial artery within the anatomical snuffbox or Hegu point,significantly reduces the incidence of radial artery occlusion compared with conventional transradial access and shortens compression time to 2-3 hours after procedure.Clinical evidence confirms the efficacy of dTRA in facilitating complex percutaneous coronary interventions,including stenting of left main coronary artery bifurcation lesions and recanalization of chronic total occlusions.Its application has progressively expanded to neurointerventional procedures(cerebral angiography),tumor embolization(transarterial chemoembolization for hepatocellular carcinoma),and peripheral vascular interventions.Despite increasing clinical adoption,dTRA still faces challenges,including a long learning curve and relatively lower initial puncture success rates.Combined with the current paucity of robust evidence-based data,the viability of dTRA as a routine interventional access route remains a subject of debate.This article systematically reviews the anatomical landmarks,clinical advantages,limitations,and multidisciplinary applications of dTRA.This article aims to provide practical guidance for interventionalists and promote the standardization of this technique in daily clinical practice.

Distal transradial access(dTRA)was first applied in coronary heart disease intervention by Kiemeneij in 2017,dTRA has become an important technological advancement of coronary interventional therapy.This approach,performed by puncturing the distal radial artery within the anatomical snuffbox or Hegu point,significantly reduces the incidence of radial artery occlusion compared with conventional transradial access and shortens compression time to 2-3 hours after procedure.Clinical evidence confirms the efficacy of dTRA in facilitating complex percutaneous coronary interventions,including stenting of left main coronary artery bifurcation lesions and recanalization of chronic total occlusions.Its application has progressively expanded to neurointerventional procedures(cerebral angiography),tumor embolization(transarterial chemoembolization for hepatocellular carcinoma),and peripheral vascular interventions.Despite increasing clinical adoption,dTRA still faces challenges,including a long learning curve and relatively lower initial puncture success rates.Combined with the current paucity of robust evidence-based data,the viability of dTRA as a routine interventional access route remains a subject of debate.This article systematically reviews the anatomical landmarks,clinical advantages,limitations,and multidisciplinary applications of dTRA.This article aims to provide practical guidance for interventionalists and promote the standardization of this technique in daily clinical practice.

Goniotomy (GT) is a safe and effective type of minimally invasive glaucoma surgery (MIGS) extensively utilized in China.It is particularly suited for treating primary open-angle glaucoma and advanced primary angle-closure glaucoma.Although GT is generally safe, hyphema and postoperative intraocular pressure (IOP) spikes remain common complications after GT.Currently, there is no standardized protocol for managing these issues, which can impact clinicians' assessment of surgical outcomes and potentially affect the prognosis.Therefore, it is crucial to establish comprehensive and detailed management protocols for perioperative hyphema and IOP spike following GT.This will guide clinical practitioners in managing complications appropriately and systematically, thereby promoting the further development and refinement of MIGS.To address these concerns, several domestic glaucoma treatment experts along with members of the Glaucoma Society of Ophthalmology, Guangdong Medical Association reviewed existing literature and held recommendation meetings to develop a guideline for managing perioperative hyphema and IOP spikes following GT.It includes defining perioperative hyphema in GT, discussing its high-risk factors and outcomes, exploring methods for prevention prior to surgery and techniques to reduce bleeding during the procedure, and managing postoperative hemorrhage.Additionally, it covers defining IOP spikes after GT, investigates their causes and contributing factors, and outlines management strategies and anticipated outcomes to provide a valuable resource for clinicians.

With high incidence, high disability rate and serious impact on patients' quality of life, advanced primary angle-closure glaucoma (PACG) with cataract is a severe eye disease in China and its main treatment method is surgery.Phacoemulsification cataract extraction and intraocular lens implantation (PEI) combined with trabeculectomy has been used to treat advanced PACG with cataract, but there are many postoperative complications.Recently, combined PEI, goniosynechialysis (GSL) and goniotomy (GT) has been proven to be a safe and effective surgical treatment for advanced PACG with cataract.At present, the combined PEI+ GSL+ GT surgery has been widely applied, but there is still a lack of uniform standards for its clinical application.Therefore, it is necessary to formulate a detailed and complete recommendation of the surgical procedures for PEI+ GSL+ GT, to standardize its application in clinical practice.Based on the analysis of the existing problems, some glaucoma experts in China have formulated detailed and complete operation norms, as well as expert recommendations on surgical indications and contraindications, anesthetic methods, surgical techniques and perioperative medication to standardize the application process of PEI+ GSL+ GT in clinical practice.

Minimally invasive glaucoma surgery (MIGS) has been successfully incorporated in clinical practice in recent years.Increasing evidence shows that MIGS also plays an increasingly important role in the treatment of primary angle-closure glaucoma (PACG). Surgical peripheral iridectomy (SPI) plus goniosynechialysis (GSL) and goniotomy (GT), as a new procedure, has been proven to be safe and effective in the treatment of PACG with no or mild cataracts.However, there is still a lack of uniform standards for indications, surgical procedures, and perioperative medication.Therefore, it is necessary to formulate a detailed and complete expert recommendations on surgical procedures to standardize the application of combined SPI+ GSL+ GT in the treatment.Some experts in glaucoma treatment and management in China have discussed the current problems and developing trend in MIGS for PACG and put forward the expert recommendations of standard process, including indications, anaesthetic methods, surgical site, operating procedure and rational administration of drugs in perioperation, in order to standardize the medical process of clinicians using combined SPI+ GSL+ GT in PACG treatment and lay a foundation for better evaluation of the efficacy of MIGS.

Objective:To evaluate the preliminary effectiveness and safety of surgical peripheral iridectomy (SPI) combined with goniosynechilysis (GSL) and goniotomy (GT) in the treatment of advanced primary angle-closure glaucoma (PACG).Methods:A multicenter observational case series study was performed.Thirty-five eyes of 27 patients with advanced PACG, who underwent SPI+ GSL+ GT with a follow-up of at least 6 months, were included from August 2021 to January 2022 at Zhongshan Ophthalmic Center, Handan City Eye Hospital, Shijiazhuang People's Hospital, West China Hospital of Sichuan University, and the Third Affiliated Hospital of Chongqing Medical University.The mean follow-up time was 9(7, 10) months.Pre- and post-operative best corrected visual acuity (BCVA) and intraocular pressure (IOP) were measured with an ETDRS chart and a Goldmann applanation tonometer, respectively.The number of anti-glaucoma medications applied before and after surgery was recorded, and the complications after surgery were analyzed.Success rate of surgery was calculated.Complete surgical success was defined as an IOP of 5-18 mmHg (1 mmHg=0.133 kPa) and 20% reduction from baseline without anti-glaucoma medication or reoperation.Qualified success was defined as achieving criterion of complete success under anti-glaucoma medications.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Zhongshan Ophthalmic Center, Sun Yat-sen University (No.2021KYPJ177). Written informed consent was obtained from each subject.Results:The mean preoperative IOP was (30.83±8.87)mmHg, which was significantly decreased to (15.69±3.70)mmHg at 6 months after the surgery ( t=8.588, P<0.001), with a 44.00% (34.78%, 60.00%) decline of 13.00(8.00, 21.00)mmHg.The median number of anti-glaucoma medications was significantly reduced from 2(0, 3) preoperatively to 0 (0, 1) postoperatively ( Z=-3.659, P<0.001). The mean preoperative and postoperative 6-month BCVA were 0.80(0.63, 1.00) and 0.80(0.60, 1.00), respectively, showing no significant difference ( Z=-0.283, P=0.777). Complete surgical success rate was 62.86%(22/35), and the qualified success rate was 91.43%(32/35). Surgical complications mainly included hyphema (6/35), IOP spike (3/35), and shallow anterior chamber (4/35). There was no vision-threatening complication. Conclusions:SPI+ GSL+ GT is preliminarily effective and safe in the treatment of advanced PACG, which provides a new option for PACG.

Minimally invasive glaucoma surgery (MIGS) has been successfully incorporated in clinical practice in recent years.Increasing evidence shows that it also plays an important role in the treatment of primary angle-closure glaucoma (PACG). Goniosynechialysis (GSL) is a key surgical procedure used by MIGS for PACG, which involves separating the peripheral anterior synechia and reopening the angle.However, there is still a lack of uniform standards for indications and surgical procedures.Therefore, it is necessary to formulate a detailed and complete expert consensus on surgical procedures to standardize the application of clinicians in the treatment.

Objective:To explore the clinical efficacy of Masquelet technique combined with tissue flap transfer in the treatment of infectious composite bone and soft tissue defects in the early and middle stages after internal fixation for tibial fractures.Methods:From October 2017 to November 2020, 12 patients (13 tibial fractures) with infectious bone and soft tissue defects in the early and middle stages after internal fixation were treated in the Department of Orthopaedics, 988th Hospital of the Joint Logistics Support Force of CPLA by two-phased surgery with retaining internal fixation. Phase I procedures were thoroughly removal of the infected lesions and failed screws, preserving internal implants as many as possible, implantation of absorbable calcium sulphate and an antibiotics blended string of beads into the distal and proximal medullary cavity of the fractured bones, filling the bone defect and wrapping the internal implants with antibiotics loaded bone cement. The size of defects was 3.5 cm × 5.0 cm-7.5 cm × 14.5 cm, and the flaps for wound coverage sized 4.0 cm × 5.5 cm-8.0 cm × 15.0 cm. As for the repair of donor site, 8 limbs were sutured directly, 5 limbs could not be closed completely, and the remaining wounds were covered by skin grafting after suture. Based on well control of infection and stable clinical signs, fillings of bone cement were then removed in Phase II surgery, or 6-9 weeks after primary surgery. Autologous cancellous bone pieces or composite allogeneic bone were fully implanted around the induction membrane formed by Masquelet technique, and auxiliary steel plates were implanted for internal fixation of unstable fractures. After discharge, the patients visited the outpatient clinic regularly, and combined with Wechat follow-up. The texture, colour and bone healing were observed. At the last follow-up, the function of the affected limbs were assessed according to Johner-Wruhs evaluation standard.Results:After Phase I surgery, 13 flaps survived smoothly without vascular compromise. The wounds healed in Phase I. Two patients (2 sides) had recurrent infections. Re-debridement was performed and external fixation was applied after removal of internal fixation. After Phase II surgery, all patients were included in 12-26 months of follow-up, with an average of 18 months. Thirteen lower leg fractures healed well, and the time of bone healing was 16-25 (average 19.5) weeks. The Johner Wruhs criteria was used in evaluation of the function of affected limbs, and it was found that 6 patients were in excellent, 5 in good and 2 in fair.Conclusion:It is feasible while preserving the internal implants, to use membrane induction technique (Masquelet technique) combined with flap transfer, together with the absorbable calcium sulphate antibiotic sustained-release beads as a carrier in the phased treatment of infectious bone defects and bone exposure in the early and middle stages after the surgery of tibial internal fixation. It also gives a higher rate of excellence in surgical outcome. This study explores a treatment procedure for traumatic bone infection combined with composite soft tissue defects.