As the need for antibody production rises, there is an urgent need to lower the costs and enhance the efficiency of the separation process. Currently, the chromatographic media used for antibody separation and purification often focus on individual properties of antibodies, such as affinity, hydrophobicity, and charge, leading to issues like low purification efficiency or inadequate adsorption capacity. To address this, an electrostatically coupled polypeptide affinity medium (FD7-3, 5-diaminobenzoic acid n-sepharose, FD7-DA-Sepharose) was developed for rapid purification of antibodies from cell culture supernatant. This medium utilized 3, 5-diaminobenzoic acid as a spacer to attach the heptapeptide-affinity ligand (FYEILHD, FD7) to agarose microspheres. Antibodies could be adsorbed through charge interactions with the carboxyl functional group of the FD7-DA-Sepharose spacer, while FD7 enhanced electrostatic coupling and affinity adsorption through synergistic effects, significantly increasing the adsorption capacity while maintaining the affinity and specificity. The influences of pH and ionic strength on adsorption capacity were investigated with human immunoglobulin as a model protein. The static adsorption capacity (Q_m) of FD7-DA-Sepharose in the solution of pH 6.0 reached 67.73 mg/mL, representing a 52.68% increase compared with that (44.36 mg/mL) of the commercial Protein A affinity medium. Furthermore, the elution conditions for FD7-DA- Sepharose were mild (20 mmol/L PB, 0.5 mol/L NaCl, pH 6.0), in contrast to the harsh acidic elution (pH 2.7-3.6) typically associated with Protein A, which can damage antibody integrity. The FD7-DA-Sepharose medium was then employed to purify antibodies from cell culture supernatant, achieving the yield of 94.8% and the purity of 98.4%. The secondary structure of the purified antibody was determined by circular dichroism spectroscopy. The results demonstrated that FD7-DA-Sepharose enabled efficient purification of antibodies from cell culture supernatant, which provided a cost-effective solution (approximately one-third the price of commercial Protein A affinity medium) with gentle elution conditions that preserve the natural conformation of antibodies. This approach paves a novel, economical, and efficient way for the separation and purification of antibodies from cell culture supernatant.
Chromatography, Affinity/methods* ; Static Electricity ; Humans ; Sepharose/analogs & derivatives* ; Peptides/chemistry* ; Adsorption ; Antibodies/isolation & purification*

Objective:To investigate the effect of β-elemene on mitochondrial structure and function of the A549 cells of non-small cell lung cancer(NSCLC),and to elucidate the mechanism of β-elemene in the treatment of NSCLC.Methods:The A549 cells at logarithmic growth stage were divided into blank control group(0 mng·L-1 β-elemene),low,medium and high doses of β-elemene groups(10,25 and 50 mg·L-1),and solvent control group(0.5％ethanol in equal volume).After treatment for 24 h,the cell activities in various groups were detected by MTT assay;the morphology changes of mitochondria in the cells in various groups was observed by transmission electron microscope;the levels of adenosine 5′-triphosphate(ATP)in the cells in various groups were detected by colorimetry;the mitochondrial membrane potential of the A549 cells in various groups were detected by JC-1 flow cytometry;mitochondrial membrane permeability transfer hole assay was used to detect the mitochondrial membrane permeabilities of the cells in various groups.Results:The MTT results showed that compared with blank control group,the cell activities in low,medium and high doses of β-elemene groups were decreased gradually(P＜0.05),while the cell activity in solvent control group had no significant change,and the difference was not significant(P＞0.05).The transmission electron microscope results showed that compared with blank control group,the mitochondria of A549 cells in low,medium and high doses ofβ-elemene groups showed swelling,vacuolation,disordered arrangement and dissolution,while the mitochondrial morphology of the A549 cells in solvent control group had no significant changes.The colorimetric method results showed that compared with blank control group,the ATP levels in the A549 cells in low,medium and high dose β-elemene groups were gradually decreased(P＜0.05),while the ATP level in the A549 cells in solvent control group had no significant change,and the difference was not significant(P＞0.05).The JC-1 flow cytometry method results showed that compared with blank control group,the mitochondrial membrane potential of the A549 cells in low,medium and high doses ofβ-elemene groups were decreased,and the percentages of the cells in Q2-4 region were increased(P＜0.05);the percentage of the A549 cells in the Q2-4 region in solvent control group had no significant change.The results of mitochondrial membrane permeability transfer hole experiment showed that compared with blank control group,the mitochondrial membrane permeabilities of the A549 cells in low,medium and high doses of β-elemene groups were increased,and the percentages of the cells in M4 region were increased(P＜0.05);the mitochondrial membrane permeability of the A549 cells and the percentage of the M4 cells in solvent control group had no significant changes,and the difference was not significant(P＞0.05).Conclusion:β-elemene can inhibit the proliferation of the A549 cells,and the mechanism may be that the mitochondrial structure of A549 cells is damaged by reducing the level of ATP and mitochondrial membrane potential,changing the mitochondrial morphology and increasing the mitochondrial membrane permeability.

Objective:To investigate the clinical characteristics and prognosis of bloodstream infection caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) in preterm infants, and to provide basis for early clinical diagnosis and infection control. Methods:The clinical data of infants with CRKP bloodstream infection admitted to the Preterm Infants Ward of Children′s Hospital Affiliated to Zhengzhou University from January 2015 to December 2022 were retrospectively analyzed. The risk factors for death in preterm infants caused by CRKP bloodstream infection were explored through multivariate logistic regression analysis, and the receiver operating characteristic (ROC) curve was used to analyze the clinical value of each factor on evaluating prognosis. The area under curves (AUC) of each factor in different ROC curve were compared by Delong′s test.Results:A total of 96 preterm infants with CRKP bloodstream infection were included, including 70 in the survival group and 26 in the death group. The first onset symptoms of CRKP bloodstream infection in preterm infants were persistent tachycardia (heart rate>180 per minute) (69 cases, 71.9%), fever (61 cases, 63.5%), and apnea (59 cases, 61.5%). There were 88(91.7%) cases of infection combined with septic shock, and 91(94.8%) cases required vasoactive drug support. Multivariate logistic regression analysis showed that the maximum vasoactive-inotropic score (VIS) within 48 hours of onset (odds ratio ( OR)=1.058, 95% confidence interval (95% CI) 1.022 to 1.095, P=0.001), concurrent purulent meningitis ( OR=8.029, 95% CI 1.344 to 47.972, P=0.022), and concurrent necrotizing enterocolitis (NEC) ( OR=10.881, 95% CI 1.566 to 75.580, P=0.016) were independent risk factors for death in preterm infants with CRKP bloodstream infection. The ROC curve showed that the AUCs for evaluating the prognosis of preterm infants with NEC and purulent meningitis were 0.784 and 0.711, respectively. The AUC for evaluating the prognosis of preterm infants with a maximum VIS ≥52.5 points within 48 hours of onset was 0.840, and the AUC for combining the three factors was 0.931. Compared with NEC and purulent meningitis, the AUC for combining factors was higher, the differences were statistically significant ( P=0.002, P<0.001). Conclusions:Preterm infants with CRKP bloodstream infection who have a maximum VIS ≥52.5 points within 48 hours of onset, with NEC and purulent meningitis have a higher risk of death.

Objective To provide evidence-based references for the prevention of surgical site infection(SSI)by sum-marizing the best evidence for the prevention of SSI in adult inpatients.Methods The'6S'evidence resource pyramid model was used to systematically search the related evidence in domestic and foreign databases,guideline websites,and academic socie-ty websites from the inception of the database to September 30,2023.Four researchers evaluated the quality of the included guidelines,and two researchers independently evaluated the quality of other types of literature and rated the level of evidence.Results A total of 12 articles were included,including 6 clinical decision making and 6 clinical guidelines.Thirty best items of the evidence were summarized from 7 aspects:diagnosis,clinical symptoms,influencing factors,patient prevention strategies,preventive strategies for medical staff,intraoperative and postoperative treatment,and consultation and education.Conclusion Clinical staff should develop a standardized management plan for infection prevention based on corresponding evidence to reduce the incidence of SSI instead of taking a single measurement.Moreover,they need to formulate a standardized work process for preventing SSI based on the clinical practice and patients'preference.

OBJECTIVE: To investigate the effect of improved nursing strategy on prognosis of older immunosuppressed patients with pneumonia and sepsis. METHODS: A prospective study was conducted. The older immunosuppressed patients with pneumonia and sepsis admitted to the department of intensive care medicine and emergency intensive care unit (ICU) of Peking Union Medical College Hospital from January 2017 to July 2022 were enrolled. In the first stage (from January 2017 to December 2019), patients received the original nursing strategy (original nursing strategy group), including: (1) nurses were randomly assigned; (2) routine terminal cleaning; (3) ICU environmental cleaning twice a day; (4) oral care was performed with chlorhexidine twice a day; (5) original lung physiotherapy [head of bed elevated at 30 degree angle-45 degree angle, maintaining a Richmond agitation-sedation scale (RASS) -2 to 1, sputum aspiration as needed]. After 1 month of learning and training of the modified nursing treatment strategy for nurses and related medical staff, the patients in the second stage (from February 2020 to July 2022) received the improved nursing strategy (improved nursing strategy group). The improved nursing strategy improved the hospital infection prevention and control strategy and lung physical therapy strategy on the basis of the original nursing strategy, including: (1) nurses were fixed assigned; (2) patients were placed in a private room; (3) enhanced terminal cleaning; (4) ICU environmental cleaning four times a day; (5) education and training in hand hygiene among health care workers was improved; (6) bathing with 2% chlorhexidinegluconate was performed once daily; (7) oral care with a combination of chlorhexidine and colistin was provided every 6 hours; (8) surveillance of colonization was conducted; (9) improved lung physiotherapy (on the basis of the original lung physiotherapy, delirium score was assessed to guide early mobilization of the patients; airway drainage was enhanced, the degree of airway humidification was adjusted according to the sputum properties, achieving sputum viscosity grade II; lung ultrasound was also used for lung assessment, and patients with atelectasis were placed in high lateral position and received the lung recruitment maneuver). Baseline patient information were collected, including gender, age, underlying diseases, source of admission, disease severity scores, vital signs, ventilatory parameters, blood gas analysis, life-sustaining treatments, clinical laboratory evaluation, indicators of infection and inflammation, pathogens and drug therapy. The primary outcome was 28-day mortality, and the secondary outcomes were duration of mechanical ventilation, length of ICU stay, and ICU mortality. Multivariate Logistic regression analysis was used to determine the risk factors for 28-day death in older immunosuppressed patients with pneumonia and sepsis. RESULTS: Finally, 550 patients were enrolled, including 199 patients in the original nursing strategy group and 351 patients in the improved nursing strategy group. No significant differences were found in gender, age, underlying diseases, source of admission, disease severity scores, vital signs, ventilatory parameters, blood gas analysis, life-sustaining treatments, clinical laboratory evaluation, indicators of infection and inflammation, coexisting pathogens or drug therapy between the two groups. Compared with patients in the original nursing strategy group, those in the improved nursing strategy group had significantly fewer duration of mechanical ventilation and length of ICU stay [duration of mechanical ventilation (days): 5 (4, 7) vs. 5 (4, 9), length of ICU stay (days): 11 (6, 17) vs. 12 (6, 23), both P < 0.01], and lower ICU mortality and 28-day mortality [ICU mortality: 23.9% (84/351) vs. 32.7% (65/199), 28-day mortality: 23.1% (81/351) vs. 33.7% (67/199), both P < 0.05]. Multivariate Logistic regression analysis showed that the improved nursing strategy acted as an independent protective factor in 28-day death of older immunosuppressed patients with pneumonia and sepsis [odds ratio (OR) = 0.543, 95% confidence interval (95%CI) was 0.334-0.885, P = 0.014]. CONCLUSIONS Improved nursing strategy shortened the duration of mechanical ventilation and the length of ICU stay, and decreased ICU mortality and 28-day mortality in older immunosuppressed patients with pneumonia and sepsis, significantly improving the short-term prognosis of such patients.
Humans ; Aged ; Prospective Studies ; Chlorhexidine/therapeutic use* ; Intensive Care Units ; Pneumonia ; Prognosis ; Sepsis/therapy* ; Inflammation

In recent years, clinical applications of robotic thyroid surgery have been gradually promoted with the continuous improvement of the da Vinci robotic surgical system. Unlike traditional open surgery, robotic thyroid surgery mainly adopts remote access, which has many advantages, such as magnified high-definition 3D view and hand vibration stabilization. The rates and causes of postoperative complications differ due to different approaches, view angles, and operation sequences. This paper presents the literature on both transoral and bilateral areolar axillary approaches in robotic thyroid surgery, focusing on five common complications under both approaches, including laryngeal recurrent nerve injury and hypoparathyroidism, to provide theoretical support for the standardization of robotic thyroid surgery.

Thyroid cancer is the most common malignant tumor of the endocrine system, and its incidence is increasing year by year showing younger trend. The pathological types of thyroid cancer are papillary carcinoma, follicular carcinoma, medullary carcinoma and undifferentiated carcinoma. Medullary thyroid carcinoma (MTC) and papillary thyroid carcinoma (PTC) are the two common types of thyroid cancer. Since MTC and PTC have different biological origins, invasiveness and prognosis. Therefore, the occurrence of both medullary carcinoma and papillary carcinoma is very rare and rarely reported in China and abroad and the treatment is challenging. In this paper, we review the pathogenesis, ancillary tests, diagnosis and treatment of this disease, with the aim of providing reference for clinical treatment of patients with medullary thyroid carcinoma complicated by papillary carcinoma.

Objective:To summarize the best evidence of intracranial hypertension nursing for adult patients with severe brain injury, and to provide reference for clinical nursing practice.Methods:According to the evidence-based methodology, a systematic search of Chinese and English literature on intracranial hypertension nursing of adult patients with severe brain injury was conducted in domestic and foreign databases such as CNKI, Wanfang, PubMed, Cochrane Library and Cinahl Plus and so on, as well as related guide websites and professional association websites from the establishment of database to August 2022. Two researchers independently evaluated literature quality and screened evidence, and then the project team summarized and concluded the evidence.Results:A total of 6 009 articles were obtained through preliminary search, and 33 articles were included after screening, including 13 guidelines, 1 systematic review, 17 expert consensus, 1 evidence summary, and 1 meta-analysis. In total, 33 pieces of best evidence were obtained from 8 dimensions, including intracranial pressure related threshold, assessment and monitoring, respiratory care, circulation care, analgesic and sedative care, temperature care, nutrition care and cerebrospinal fluid care.Conclusions:This study summarizes the evidence-based basis of intracranial hypertension nursing in adult patients with severe brain injury, which provides a basis for the standardized construction of clinical nursing strategies and empirical research.

Owing to complex surgical planning and procedures, orthognathic surgery requires surgeons to possess a high level of clinical expertise. However, traditional teaching methods are difficult to meet the requirements for complex training on orthognathic surgical planning and operation. Virtual reality (VR) technology provides a new solution, but it is limited by problems like imprecise modeling, complex surgical design, single-simulated surgical contents, inability to integrate real time and simulation, and a lack of automatic evaluation function. Artificial intelligence (AI) is potent in data mining and image processing, which can address the limitations of traditional VR teaching, bringing multiple potential for clinical and educational application in orthognathic surgery. With research in personalized model construction, intelligent diagnostics, post-operative prediction, virtual surgical simulation, and personalized feedback and guidance, the fusion of "VR plus AI" holds promise for advancing orthognathic surgical teaching in a more precise and efficient direction, promoting the integration of medicine and technology, and speeding up the transformation of clinical applications.

OBJECTIVES: Safe and effective anticoagulation is essential for hemodialysis patients who are at high risk of bleeding. The purpose of this trial is to evaluate the effectiveness and safety of two-stage regional citrate anticoagulation (RCA) combined with sequential anticoagulation and standard calcium-containing dialysate in intermittent hemodialysis (IHD) treatment. METHODS: Patients at high risk of bleeding who underwent IHD from September 2019 to May 2021 were prospectively enrolled in 13 blood purification centers of nephrology departments, and were randomly divided into RCA group and saline flushing group. In the RCA group, 0.04 g/mL sodium citrate was infused from the start of the dialysis line during blood draining and at the venous expansion chamber. The sodium citrate was stopped after 3 h of dialysis, which was changed to sequential dialysis without anticoagulant. The hazard ratios for coagulation were according to baseline. RESULTS: A total of 159 patients and 208 sessions were enrolled, including RCA group (80 patients, 110 sessions) and saline flushing group (79 patients, 98 sessions). The incidence of severe coagulation events of extracorporeal circulation in the RCA group was significantly lower than that in the saline flushing group (3.64% vs. 20.41%, P<0.001). The survival time of the filter pipeline in the RCA group was significantly longer than that in the saline flushing group ((238.34±9.33) min vs. (221.73±34.10) min, P<0.001). The urea clearance index (Kt/V) in the RCA group was similar to that in the saline flushing group with no statistically significant difference (1.12±0.34 vs. 1.08±0.34, P=0.41). CONCLUSIONS Compared with saline flushing, the two-stage RCA combined with a sequential anticoagulation strategy significantly reduced extracorporeal circulation clotting events and prolonged the dialysis time without serious adverse events.
Humans ; Citric Acid/adverse effects* ; Prospective Studies ; Sodium Citrate ; Hemorrhage/chemically induced* ; Citrates/adverse effects* ; Anticoagulants/adverse effects* ; Renal Dialysis/adverse effects*