Objective To prepare humanized monoclonal antibodies(Mabs)targeting Acinetobacter baumannii(Ab)based on single memory B cell sequencing technology,construct the immune repertoire of the core protein of Ab,hemolysin-coregulated protein(Hcp),and express its Mabs with binding activity.Methods E.coli BL21 harboring the recombinant plasmid pGEX-6p-1-Hcp was constructed.Hcp protein was obtained using protein expression and affinity chromatography.Female SPF BALB/c mice(6～8 weeks old,weighing 18～20 g)were immunized intramuscularly with antigen Hcp to generate specific memory B cells.Single antigen-specific memory B cells were sorted using flow cytometry.The immune repertoire of Hcp was constructed using single-cell sequencing technology,and bioinformatics analysis was performed on the sequencing results.Mabs were obtained using antibody humanization techniques.The in vitro binding activity of the antibodies was detected by ELISA.Results The target protein Hcp with a purity>95%was obtained after expression and purification.The immune repertoire of Hcp was successfully constructed,and the results of BCR clonotype identification and analysis,CDR3 region characteristic analysis,and V-J gene pairing characteristic analysis were achieved.Antibody humanization got 7 Mabs,that is,IgG1-1,IgG1-2,IgG2-1,IgG2-2,IgG3-1,IgG4-1 and IgG4-2.ELISA results showed IgG1-1,IgG3-1,IgG4-1,and IgG4-2 had an antibody binding titer of 1∶1 280,IgG2-2 of 1∶10 240,IgG2-1 of 1∶5 120,and IgG1-2 of 1∶160.Conclusion Single-cell sequencing technology enables rapid,accurate,and efficient construction of an Hcp protein immune repertoire containing extensive antibody information.Utilizing this immune repertoire allows for the expression of Mabs with binding activity.

Objective:To explore the effect of enhanced recovery after surgery (ERAS) protocols in patients undergoing laparoscopic radical cystectomy (LRC) and intracorporeal urinary diversion (ICUD).Methods:A total of 83 patients who received LRC+ ICUD in Beijing Chaoyang Hospital from March 2014 to September 2020, were divided into 2 groups based on different perioperative management, including 29 ERAS cases and 54 conventional recovery after surgery (CRAS) cases. The ERAS group included 26 males and 3 females , with an average age of (62.07 ± 9.26) years. There were 26 patients with ASA class Ⅰ-Ⅱ, 3 patients with ASA class Ⅲ, 4 patients received neoadjuvant chemotherapy, and 7 patients had a history of abdominal surgery in ERAS group. The CRAS group included 44 males and 10 females , with an average age of (61.59 ± 10.16) years. There were 50 patients with ASA class Ⅰ-Ⅱ, 4 patients with ASA class Ⅲ, 9 patients received neoadjuvant chemotherapy, and 10 patients had a history of abdominal surgery in CRAS group. There were no statistically significant differences in the baseline characteristics between the two groups. The patients in both groups underwent LRC+ ICUD procedures. The perioperative results and complications between the two groups were compared.Results:In the ERAS group, there were 20 patients who underwent Bricker ileal conduit surgery and 9 patients who underwent Studer orthotopic ileal neobladder surgery. Pathological staging included 3, 3, 7, 7, 5 and 4 cases in stage T a, T is, T 1, T 2, T 3 and T 4a, respectively. There were 23, 2, 3 and 1 patient with pathological stage N 0, N 1, N 2 and N 3, respectively. Pathological diagnosis included 3 cases of low-grade urothelial carcinoma, 24 cases of high-grade urothelial carcinoma, and 2 cases of other histological subtypes. In the CRAS group, there were 31 patients who underwent Bricker ileal conduit surgery and 23 patients who underwent Studer orthotopic ileal neobladder surgery. Pathological staging included 5, 3, 12, 9, 15 and 10 patients in stage T a, T is, T 1, T 2, T 3 and T 4a, respectively. There were 35, 6, 7 and 6 patients with pathological stage N 0, N 1, N 2, and N 3, respectively. Pathological diagnoses included 6 cases of low-grade urothelial carcinoma, 45 cases of high-grade urothelial carcinoma, and 3 cases of other histological subtypes. There were no statistically significant differences ( P>0.05) in surgical methods, pathological staging, or pathological types between the ERAS and CRAS groups. ERAS group presented less albumin loss [(25.73±8.63)% vs. (32.63±9.05)%, P=0.001], shorter hospital stay [9(7, 13)d vs. 12(9, 16)d, P=0.006], less 30-day overall complications [55.2% (16/29) vs. 83.3% (45/54), P=0.009]. In multivariable analysis, maximum albumin loss≥20% was independently associated with 30-day minor complications ( P=0.049), and maximum albumin loss ≥25% was independently associated with hospital of stay≥10 days ( P=0.038), respectively. Conclusions:For patients who received LRC+ ICUD, ERAS was associated with reduced perioperative albumin loss, shorter length of stay, less 30-day complications, accelerated recovery time, improved clinical outcome and less albumin injection.

Objective:To explore the early learning curve of OrthoPilot navigation assisted total knee arthroplasty (TKA).Methods:Data of 40 consecutive cases of OrthoPilot navigation assisted TKA completed by the same surgical team in our department were retrospectively analyzed. According to the operation order, 40 cases were divided into the original phase group (the first 20 cases) and the subsequent phase group (the second 20 cases). In original phase group, the average age was 69.85±6.86 years with mean body mass index 24.10±2.88 kg/m 2, preoperative HSS score 48.80±5.33, preoperative knee ROM 87.05°±11.02° and preoperative alignment deviation of 7.40°±5.59°. In subsequent phase group, the average age was 66.65±7.92 years with mean body mass index 22.85±3.15 kg/m 2, preoperative HSS score 49.00±5.47, preoperative knee ROM 85.80°±11.65° and preoperative alignment deviation of 8.22°±5.21°. Perioperative data such as operative duration, incision length, hemoglobin drop and postoperative hospital stay, radiographic outcomes including hip-knee-ankle angle (HKAA), mechanical lateral distal femoral angle (mLDFA), mechanical medial proximal tibial angle (mMPTA), sagittal femoral component angle (sFCA), sagittal tibial component angle (sTCA), joint line convergence angle (JLCA), and functional scores were compared between the two groups. Results:All 40 cases were followed up for 24-33 months (mean, 27.38± 2.73 months). No severe postoperative complications such as infection and loosening occurred during the follow-up. The mean operative duration was 112.35±25.49 min in original phase group versus 82.10±10.96 min in subsequent phase group ( P< 0.05). The durations of tibial cutting was 11.95±3.27 min in original phase group versus 7.35±2.23 min in subsequent phase group ( P< 0.05); the femoral planning + cutting time was 20.95±6.91 min in original phase group versus 16.60±4.78 min in subsequent phase group, and trial + prosthesis implantation time was 39.65±7.72 min in original phase group versus 25.10±5.72 min in subsequent phase group,which was significantly higher in original phase group. There was no significant difference in other perioperative data such as incision length, hemoglobin drop and postoperative hospital stay between the two groups. As for radiographic outcomes, there was no statistical difference between the two groups in the postoperative angular deviation of HKAA (0.70°±0.80° vs. 0.80°±1.06°), mLDFA (0.89°±0.91° vs. 1.00°±0.86°), mMPTA (0.77°±0.53° vs. 0.76°±1.03°), sFCA (0.73°±0.48° vs. 0.87°±1.06°), sTCA (0.95°±0.58° vs. 1.16°±1.14°) and JLCA (0.27°±0.25° vs. 0.39°±0.18°). In original phase group, the HSS scores preoperative and 3 days postoperative were 48.80±5.33 and 60.05±5.10 respectively, and those in subsequent phase were 49.00±5.47 and 60.75±4.47 respectively, and both groups showed satisfactory functional recovery. There was no significant difference in HSS scores at all follow-up time points between two phases, as well as ROM (113.20°±9.82° vs. 113.50°±12.44°) and FJS-12 scores (78.00°±10.98° vs. 76.65°±10.29°) at 2 years postoperatively. Conclusion:In this study, we described a time-related early learning curve for OrthoPilot navigation-assisted TKA, in which the operative duration tended to be shorter after the first 20 cases. However, benefiting from good operative accuracy and repeatability, satisfactory radiographic and functional outcomes can be obtained in early stage of the learning curve.

Objective:To analyze the learning effect of laparoscopic radical cystectomy(LRC)+ modified ileal conduit(MIC).Methods:From 2014 to 2019, 42 patients underwent MIC and their clinical data was retrospectively analyzed. 34 operations were performed by surgeon 1 and 8 operations by surgeon 2. We divided the 34 patients of surgeon 1 into three groups according to their surgical sequence (group A, 1st to 12th; group B, 13th to 23th; group C, 24 th to 34 th), the 8 cases of surgeon 2 was regarded as group D. The history of abdomen surgery in the 4 groups were 0, 1, 4, 3 cases, respectively ( P<0.05). There was no significant difference of the other baseline characteristics, such as age, BMI, American Society of Anesthesiologists. Then we compared several variables between the 4 groups like operation time, time of ileal conduit construction, blood loss, complication rate, lymph node yield, surgical margin, etc. The key steps of the MIC included isolating terminal ileum when the mesentery was transilluminated, performing end-to-end reflux ureterointestinal anastomosis after the efferent loop was fixed, closing the rent of the retroperitoneum. Results:All operations were performed intracorporeally with no transition to open surgery. The operative time for group A, B, C were 330.0(320.0, 360.0)min, 300.0(250.0, 308.0)min, 270.0(216.0, 324.0)min, respectively ( P =0.010). The time of ileal conduit construction of the 3 groups were 136.5(131.3, 147.5)min, 92.0(79.0, 119.0)min, 79.0(72.0, 115.0)min, respectively ( P <0.001). In addition, the difference of the two variables above between A and B, A and C groups separately reached statistical significance ( P<0.05), while the difference between B and C groups did not ( P>0.05). Other variables, such as blood loss, complication rate, lymph node yield, surgical margin, between the 3 groups reached no statistical significance ( P>0.05). The operative time of group D was 420.0(350.0, 450.0)min, and it reached statistical significance ( P<0.05) when compared with group A. There were no significant differences in other variables, such as blood loss, complication rate, lymph node yield, surgical margin, among the 2 groups ( P>0.05). Conclusions:The learning effect of LRC+ MIC was obvious. When surgeon volume increased, the operative time decreased significantly. Variables like estimated blood loss and complication rate of the 2 surgeons did not reached significant difference, which indicated reproductivity and safety of this procedure.

Objective To assess the value of clinical applications of whole-body diffusion-weighted imaging (WB-DWI) in diagnosing patients with malignant tumors compared with positron emission tomography (PET). Methods A total of 22 patients with highly suspected malignant tumors underwent WB-DWI after PET. The differences between the two imaging methods were compared in displaying lesions, and the correlation between ADC and SUV value was analyzed. Results More lesions were showed with WB-DWI than PET. There was no significant difference between the two methods in detecting the lesions of lung, mediastinal septum or abdomen (P>0.05), but more lesions in skeleton were showed with WB-DWI (P<0.05). No significant correlation between ADC and SUV value was found. Conclusion Compared with PET, WB-DWI can detect more tumor lesions. The sensitivity of WB-DWI in detecting metastatic tumors of bone is higher than that of PET.

OBJECTIVETo verify the linkage of the candidate regions identified in a previous study (markers D2S168, D2S151, D2S142 on chromosome 2) with hypertension in Chinese families.

METHODSA genetic linkage study focused on chromosome 2 was performed on 240 Chinese families containing 856 patients with essential hypertension. A total of 1080 individuals were genotyped using 9 highly polymorphic microsatellite markers around the candidate regions on chromosome 2 with an average spacing of 5 cM. Non-parametric linkage (NPL), parametric linkage analysis and transmission-disequilibrium test (TDT) with the GENEHUNTER software were used to assess evidence for linkage.

RESULTSLinkage of a region on chromosome 2 around D2S151 and D2S142 with hypertension was confirmed by different statistical methods (NPL, LOD score and TDT). However, the linkage of D2S168 could not be replicated in this extension study.

CONCLUSIONSThe data suggest that a region on chromosome 2 at or near the loci of D2S142 and D2S151 may harbor genes responsible for the development of essential hypertension in Chinese.

Alleles ; China ; Chromosomes, Human, Pair 2 ; genetics ; Family Health ; Female ; Gene Frequency ; Genetic Linkage ; Humans ; Hypertension ; genetics ; Linkage Disequilibrium ; Male ; Microsatellite Repeats