Objective To construct a library of human-derived anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) single-chain variable fragments (scFv) and screen for broad-spectrum neutralizing antibodies to identify candidate molecules for the development of diagnostic and therapeutic agents. Methods Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of patients who had recovered from novel coronavirus infection. Total RNA was extracted from these PBMCs and reverse transcribed into cDNA, which was used as a template for constructing a human anti-SARS-CoV-2 scFv library. Phage display technology was used to screen for scFv antibodies specific to the SARS-CoV-2 S protein. Full-length IgG antibodies were synthesized through sequence analysis and human IgG expression, and their binding capacity and neutralizing activity against SARS-CoV-2 were evaluated. Results A human-derived scFv antibody library against SARS-CoV-2 with a capacity of 1.56×10⁷ CFU was successfully constructed. Two specific scFv antibodies were screened from this library and expressed as full-length IgG antibodies (IgG-A10 and IgG-G6). IgG-A10 exhibited strong neutralizing activity against both the original SARS-CoV-2 strain (WT) and the XBB subvariant of the Omicron variant. However, the neutralizing activity of this antibody against the JN.1 sub lineage of the Omicron BA.2.86 variant was moderate. Conclusion This study has successfully constructed a human anti-SARS-CoV-2 scFv antibody library from the peripheral blood of recovered patients, and screened and expressed anti-SARS-CoV-2 IgG antibodies with neutralizing activity, laying a foundation for the prevention, diagnosis, and treatment of SARS-CoV-2 infection.
Humans ; Single-Chain Antibodies/genetics* ; SARS-CoV-2/immunology* ; COVID-19/immunology* ; Immunoglobulin G/genetics* ; Antibodies, Viral/genetics* ; Peptide Library ; Spike Glycoprotein, Coronavirus/immunology* ; Antibodies, Neutralizing/immunology* ; Leukocytes, Mononuclear/immunology* ; Broadly Neutralizing Antibodies/immunology*

Objective:To explore the correlation between serum 1, 5-anhydroglucitol (1, 5-AG) and mild cognitive impairment (MCI) in elderly patients with type 2 diabetes mellitus (T2DM).Methods:A total of 160 patients with T2DM aged 60-75 years old who visited the First Hospital of Hebei Medical University from May 2021 to July 2022 were selected. According to the Montreal cognitive assessment (MoCA), all patients were divided into T2DM with MCI group (T2DM+ MCI group, n=81) and T2DM without MCI group (T2DM group, n=79). All research subjects were tested for glycated hemoglobin A1c (HbA1c), serum 1, 5-AG, serum β-amyloid peptide 42 (Aβ42), and blood biochemical indicators.SPSS 25.0 statistical software was used for data analysis. The t test, Mann-Whitney U test and χ2 test were used to compare the two groups. Binary Logistic regression analysis was used to examine the relevant influencing factors. Results:(1) Compared with T2DM group, patients in T2DM+ MCI group had significantly higher age, systolic pressure and HbA1c(all P<0.05).The level of 1, 5-AG in T2DM+ MCI group was significantly lower than that in T2DM group( (15.65±2.56 )μg/mL, (18.17±3.72 )μg/mL, P<0.01), and the level of Aβ42 was higher than that of T2DM group (2.95 (3.36) pg/mL, 1.91 (2.48) pg/mL, P<0.05). (2) Binary Logistic regression analysis results showed that HbA1c( β=0.230, OR=1.259, 95% CI=1.010-1.568, P=0.040) and Aβ42( β=0.188, OR=1.206, 95% CI=1.033-1.409, P=0.018) were the independent risk factors for MCI in elderly patients with T2DM, while 1, 5-AG ( β=-0.240, OR=0.786, 95% CI=0.698-0.886, P<0.001) was the protective factor for MCI. Conclusion:There is a positive correlation between serum 1, 5-AG and cognitive function, and the decrease of 1, 5-AG level was associated with the increased risk of MCI in elderly patients with T2DM.

Hand osteoarthritis(HOA)is a disease of hand joint disorders,mainly manifested by hand interphalangeal joint and thumb carpal metacarpal joint pain,swelling,morning stiffness,limited movement,and even deformity,belonging to the category of TCM"bone arthralgia".The authors believe that HOA is more common with Taiyang Shaoyang disease,suitable for simultaneous treatment for Taiyang and Shaoyang,to operate the cardinal,regulate qi,blood,nutritive and defensive levels,dispel wind and cold,remove dampness and arthralgia,using modified Chaihu Guizhi Decoction,with confirmed efficacy.

Lumbar disc herniation is mainly manifested as lower back pain,numbness,weakness,and radiating pain in the lower limbs,which seriously affects the patients'work and quality of life.In clinical practice,it has been found that this disease always belongs to the category of deficiency in healthy qi and excess in pathogenic factors,often accompanied by kidney deficiency and cold dampness.Kidney deficiency is the root cause,while cold dampness is the symptoms.The two factors interact with each other and cause back pain.The treatment is based on dispersing cold and dampness,tonifying the kidneys and strengthening the waist,and the classic ancient formula Shenzhuo Powder is safe and effective.

Objective:To investigate the accuracy and feasibility of applying rapid immunohistochemistry(IHC) with CK5/6 antibodies in prostate biopsy tissues to assist frozen pathology diagnosis.Methods:The data of 41 patients who underwent prostate puncture and frozen tissue rapid IHC with CK5/6 antibody in Beijing Hospital from October 2022 to April 2023 were retrospectively analyzed. The median age of the patients was 76 (69, 79) years old, and the median PSA value was 12.37 (7.07, 26.17) ng/ml.The Prostate Imaging Reporting and Data System (PI-RADS) scores of the target lesions were all ≥3. The PI-RADS score of 9 patients(21.95%) was 3, 15 (36.59%) was 4, and 17(41.46%) was 5. The median diameter of the lesions in the MRI examination was 1.40 (1.09, 2.20) cm.Fourteen lesions (34.14%) were located in the migratory zone, 23 (56.10%) were located in the peripheral zone, and 4 (9.76%) involved both peripheral and migratory zone lesions. Transperineal cognitive fusion targeted combined systematic biopsy was used, and intraoperatively, 1 additional needle was taken from each of the target and non-target areas for frozen pathology section, and hematoxylin eosin(HE) staining and rapid IHC staining with CK5/6 antibody was performed, then the frozen remaining tissue was HE staining and CK5/6 IHC staining. Data such as HE and rapid IHC results of frozen pathology sections and HE and IHC results of routine sections of the frozen remaining tissues, International Society of Urological Pathology (ISUP) grading groupings(GG), and actual diagnostic results of targeted combined systematic puncture were recorded. Using the routine IHC results of the same needle tissue as the gold standard, the sensitivity and positive predictive value of applying rapid IHC frozen pathology to diagnose prostate cancer and the accuracy of its pathological GG were analyzed.Results:Among the 41 patients, a total of 35 cases were diagnosed with prostate cancer(PCa) by HE staining in frozen section of target tissue, with a positivity rate of 85.37%(35/41). Among these, there were 17 cases (48.57%) in ISUP GG 1, 8 cases (22.86%) in GG 2, 4 cases (11.43%) in GG 3, and 6 cases (17.14%) in GG 4 to 5. In addition, a total of 35 cases were diagnosed with PCa by HE staining in frozen remaining section of target tissue, with a positivity rate of 85.37%(35/41). Among these, there were 17 cases (48.57%) in ISUP GG 1, 8 cases (22.86%) in GG 2, 4 cases (11.43%) in GG 3, and 6 cases (17.14%) in GG 4 to 5. The results of rapid IHC of target tissue: 35 cases were negative for CK5/6 expression and 6 cases were positive. The results of routine IHC of target tissue: 35 cases were negative for CK5/6 expression and 6 cases were positive. The results of rapid IHC of non-target tissue: 12 cases were negative for CK5/6 expression and 29 cases were positive. The results of routine IHC of non-target tissue: 12 cases were negative for CK5/6 expression and 29 cases were positive. Thirty-five cases of target tissue rapid IHC diagnosis of PCa were in complete agreement with routine IHC diagnosis, with a false-positive rate of 0, a sensitivity of 100.00% (35/35) and a positive predictive value of 100.00% (35/35). Twelve cases of non-target tissue rapid IHC diagnosis of PCa were in complete agreement with routine IHC diagnosis, with a false-positive rate of 0, a sensitivity of 100.00% (35/35), and a positive predictive value of 100.00% (12/12).Conclusions:The preliminarily study results confirmed that the application of rapid IHC with CK5/6 antibodies in prostate biopsy tissues assisted frozen pathology diagnosis with high accuracy, but the reliability of rapid IHC technology in assisting frozen pathological diagnosis during puncture surgery still needs further validation through large sample size prospective studies.

Objective:To evaluate the effects of different subanesthetic doses of esketamine on lung injury in elderly patients undergoing robot-assisted radical prostatectomy.Methods:Ninety American Society of Anesthesiologists Physical Status classification Ⅱ or Ⅲ patients, aged 65-80 yr, with body mass index of 19-27 kg/m 2, scheduled for elective robot-assisted radical prostatectomy under general anesthesia, identified as having middle and high risk using the Assess Respiratory Risk in Surgical Patients in Catalonia, were divided into 3 groups ( n=30 each) using a random number table method: low-dose esketamine group (ES1 group), extremely low-dose esketamine group (ES2 group) and control group (C group). In ES1 group, esketamine was intravenously injected as a bolus of 0.2 mg/kg during anesthesia induction followed by an infusion of 0.125 mg·kg -1·h -1 until 30 min before the end of operation. In ES2 group, esketamine was intravenously injected as a bolus of 0.1 mg/kg during anesthesia induction followed by an infusion of 0.015 mg·kg -1·h -1 until 30 min before the end of operation. The equal volume of normal saline was given instead in C group. Radial artery blood samples were collected before anesthesia induction (T 0) and at the end of operation for determination of concentrations of Clara cell secretory protein (CC-16) and soluble form of advanced glycation end products receptor (sRAGE) in serum by enzyme-linked immunosorbent assay. The parameters of respiratory mechanics such as the driving pressure, dynamic lung compliance and mechanical power were recorded at 5 min after mechanical ventilation (T 1), and at 1 and 2 h after Trendelenburg position combined with pneumoperitoneum (T 2-3), and at 5 min before the end of operation (T 4). Blood samples were collected from the radial artery at T 0, T 1, T 3 and in the postanesthesia care unit for blood gas analysis, and the alveolar-arterial partial oxygen pressure difference and oxygenation index were recorded. The adverse reactions within 24 h after operation and the occurrence of postoperative pulmonary complications within 7 days after operation were recorded. Results:Compared with C group, the serum CC-16 and sRAGE concentrations were significantly decreased at the end of operation, the oxygenation index was increased and the alveolar-arterial partial oxygen pressure difference was decreased in the postanesthesia care unit, and the incidence of postoperative nausea reactions within 24 h after operation was decreased in ES1 and ES2 groups ( P<0.05 or 0.01). Compared with ES2 group, the serum CC-16 and sRAGE concentrations were significantly decreased at the end of operation in ES1 group ( P<0.05). There were no statistically significant differences in the driving pressure, dynamic lung compliance and mechanical power at T 1-4 and the incidence of postoperative pulmonary complications within 7 days after surgery among the three groups ( P>0.05). Conclusions:Esketamine given as a subanesthetic bolus of 0.2 mg/kg during anesthesia induction followed by an infusion of 0.125 mg·kg -1·h -1 can alleviate lung injury in elderly patients undergoing robot-assisted radical prostatectomy.

Liver transplantation(LT)is the only effective treatment for hepatopulmonary syndrome(HPS).Moreover,perioperative refractory hypoxemia(pRH)is a prevalent life-threatening condition and has extremely limited treatment options.Here,we report three patients with HPS who experienced pRH after LT and were consecutively treated with different salvage therapies,ephedrine inhalation,intravenous use of methylene blue with nitric oxide(NO)inhalation,and NO inhalation alone.The results showed that unresolved severe hypoxia may induce fatal morbidity such as early biliary leakage and acute kidney injury.Early initiation of NO inhalation,rather than ephedrine,can significantly improve oxygenation in patients with pRH and may help prevent hypoxia-related complications.Therefore,based on the response to these exploratory salvage treatments,we further demonstrate the unique ventilation-perfusion mismatch pathophysiology in specific lung regions during pRH in HPS.We propose that early inhalation of NO is an important treatment option to rescue severe hypoxia in patients with HPS during the perioperative period of LT.

Objective:To investigate the safety and therapeutic effect of split liver transplantation (SLT) in clinical application.Methods:This is a retrospective case-series study. The clinical data of 203 consecutive SLT, 79 living donor liver transplantation (LDLT) and 1 298 whole liver transplantation (WLT) performed at the Third Affiliated Hospital of Sun Yat-sen University from July 2014 to July 2023 were retrospectively analyzed. Two hundred and three SLT liver grafts were obtained from 109 donors. One hundred and twenty-seven grafts were generated by in vitro splitting and 76 grafts were generated by in vivo splitting. There were 90 adult recipients and 113 pediatric recipients. According to time, SLT patients were divided into two groups: the early SLT group (40 cases, from July 2014 to December 2017) and the mature SLT technology group (163 cases, from January 2018 to July 2023). The survival of each group was analyzed and the main factors affecting the survival rate of SLT were analyzed. The Kaplan-Meier method and Log-rank test were used for survival analysis.Results:The cumulative survival rates at 1-, 3-, and 5-year were 74.58%, 71.47%, and 71.47% in the early SLT group, and 88.03%, 87.23%, and 87.23% in the mature SLT group, respectively. Survival rates in the mature SLT group were significantly higher than those in the early SLT group ( χ2=5.560, P=0.018). The cumulative survival rates at 1-, 3- and 5-year were 93.41%, 93.41%, 89.95% in the LDLT group and 87.38%, 81.98%, 77.04% in the WLT group, respectively. There was no significant difference among the mature SLT group, the LDLT group and the WLT group ( χ2=4.016, P=0.134). Abdominal hemorrhage, infection, primary liver graft nonfunction,and portal vein thrombosis were the main causes of early postoperative death. Conclusion:SLT can achieve results comparable to those of WLT and LDLT in mature technology liver transplant centers, but it needs to go through a certain time learning curve.

Objective:To investigate the safety and therapeutic effect of split liver transplantation (SLT) in clinical application.Methods:This is a retrospective case-series study. The clinical data of 203 consecutive SLT, 79 living donor liver transplantation (LDLT) and 1 298 whole liver transplantation (WLT) performed at the Third Affiliated Hospital of Sun Yat-sen University from July 2014 to July 2023 were retrospectively analyzed. Two hundred and three SLT liver grafts were obtained from 109 donors. One hundred and twenty-seven grafts were generated by in vitro splitting and 76 grafts were generated by in vivo splitting. There were 90 adult recipients and 113 pediatric recipients. According to time, SLT patients were divided into two groups: the early SLT group (40 cases, from July 2014 to December 2017) and the mature SLT technology group (163 cases, from January 2018 to July 2023). The survival of each group was analyzed and the main factors affecting the survival rate of SLT were analyzed. The Kaplan-Meier method and Log-rank test were used for survival analysis.Results:The cumulative survival rates at 1-, 3-, and 5-year were 74.58%, 71.47%, and 71.47% in the early SLT group, and 88.03%, 87.23%, and 87.23% in the mature SLT group, respectively. Survival rates in the mature SLT group were significantly higher than those in the early SLT group ( χ2=5.560, P=0.018). The cumulative survival rates at 1-, 3- and 5-year were 93.41%, 93.41%, 89.95% in the LDLT group and 87.38%, 81.98%, 77.04% in the WLT group, respectively. There was no significant difference among the mature SLT group, the LDLT group and the WLT group ( χ2=4.016, P=0.134). Abdominal hemorrhage, infection, primary liver graft nonfunction,and portal vein thrombosis were the main causes of early postoperative death. Conclusion:SLT can achieve results comparable to those of WLT and LDLT in mature technology liver transplant centers, but it needs to go through a certain time learning curve.

Objective To investigate the diagnosis and treatment strategy of the portal vein complications in children undergoing split liver transplantation. Methods The clinical data of 88 pediatric recipients who underwent split liver transplantation were retrospectively analyzed. Intraoperative anastomosis at the bifurcating site of the portal vein or donor iliac vein bypass anastomosis was performed depending on the internal diameter and development of the recipient's portal vein. A normalized portal venous blood stream monitoring was performed during the perioperative stage. After operation, heparin sodium was used to bridge warfarin for anticoagulation therapy. After portal vein stenosis or thrombosis was identified with enhanced CT or portography, managements including embolectomy, systemic anticoagulation, interventional thrombus removal, balloon dilatation and/or stenting were performed. Results Among the 88 recipients, a total of 10 children were diagnosed with portal vein complications, of which 4 cases were diagnosed with portal vein stenosis at 1 d, 2 months, 8 months, and 11 months after surgery, and 6 cases were diagnosed with portal vein thrombosis at intraoperative, 2 d, 3 d (n=2), 6 d, and 11 months after surgery, respectively. One patient with portal vein stenosis and one patient with portal vein thrombosis died perioperatively. The fatality related to portal vein complications was 2% (2/88). Of the remaining 8 patients, 1 underwent systemic anticoagulation, 2 underwent portal venous embolectomy, 1 underwent interventional balloon dilatation, and 4 underwent interventional balloon dilatation plus stenting. No portal venous related symptoms were detected during postoperative long term follow up, and the retested portal venous blood stream parameters were normal. Conclusions The normalized intra- and post-operative portal venous blood stream monitoring is a useful tool for the early detection of portal vein complications, the early utilization of useful managements such as intraoperative portal venous embolectomy, interventional balloon dilatation and stenting may effectively treat the portal vein complications, thus minimizing the portal vein complication related graft loss and recipient death.