Objective: To optimize the technology and process for recovering immune cells from discarded leukocyte filter through reverse flushing, and to investigate the effects of different reverse flushing solutions and physical conditions (temperature, soaking, and ultrasound) on the cell recovery rate. Additionally, to evaluate the feasibility and quality of isolating and culturing NK cells from the recovered cells. Methods: Normal saline, PBS, and PBS-EDTA were used as reverse flushing solutions to backwash the filter, and the white blood cell recovery rates were compared. The reverse flushing efficiency was optimized by adjusting the temperature of the reverse flushing solution, filter soaking treatment and ultrasonic treatment. The Ficoll separation method was employed to isolate peripheral blood mononuclear cells (PBMCs) from the elution solution and conduct in vitro expansion and culture. Lymphocyte subsets were identified and quantified using flow cytometry. Results: The physiological saline served as the backflush fluid was the most stable. The reverse temperature had a significant impact on the recovery rate of white blood cell subgroups. The recovery rates of lymphocytes and monocytes at 37℃ were significantly higher than those at 4℃ [(35.56±5.55)% vs (22.88±0.31)%, (32.42±4.82)% vs (8.00±1.6)%, t=5.581 and 5.758, respectively, both P<0.001)], and at 25℃ [(35.56±5.55)% vs (22.42±6.13)%, t=5.904, P<0.001; (32.4±4.82)% vs (21.11±1.23)%, t=2.629, P=0.030)]. The recovery rates of white blood cell subgroups after 37℃ immersion were higher than those after direct reverse flow at 37℃, but the difference was not statistically significant (P>0.05). The recovery rate of cells after ultrasonic immersion at 25℃ was further increased compared to that after immersion at 25℃, but the difference was not statistically significant (P>0.05). The purity of the NK cells obtained through sorting reached (94.2±4.5)%, and the viability was (94.8±0.9)%. After 15 days of culture, the cell count was expanded approximately 450 times, yielding more than 10 /mL highly active NK cells. Conclusion: By optimizing the backflushing process, immune cells can be efficiently recovered from discarded filters. The 37℃ backflow liquid has the best backflow effect; soaking and ultrasonic treatment can be used as auxiliary methods to improve recovery rates. The NK cells from this source have high purity, good activity, and strong expansion ability, and can be used as a reliable cell source for immunotherapy research, and in line with the circular economy concept of resource reuse.

Objective To investigate the short-term efficacy and safety of small-incision laparo-scopic radical gastrectomy in patients with advanced gastric cancer.Methods A total of 80 patients with advanced gastric cancer scheduled for radical gastrectomy were enrolled and randomly divided into control group and observation group using a random number table method,with 40 patients in each group.The control group underwent open surgery,while the observation group underwent small-inci-sion laparoscopic surgery.Clinical indicators related to the surgery were recorded in both groups.In-flammatory markers were measured and compared between the two groups before surgery and 72 hours af-ter surgery.Postoperative recovery and the incidence of complications during hospitalization were ob-served and compared between the two groups.Results There were no statistically significant differences in operative time,the number of dissected lymph nodes,distance from the proximal margin to the tumor,and distance from the distal margin to the tumor between the two groups(P＞0.05).The incision length in the observation group was shorter,intraoperative blood loss was less,hospital stay was shorter,and treatment cost was higher than those in the control group[(7.15±1.22)cm versus(14.65±2.64)cm,(177.16±28.46)mL versus(244.16±42.16)mL,(8.11±1.46)d versus(14.15±3.65)d,(5.54±0.31)×10,000 yuan versus(4.38±0.61)×10,000 yuan,P＜0.05].There were no sta-tistically significant differences in preoperative levels of C-reactive protein,interleukin-6,and tumor necrosis factor-α between the two groups(P＞0.05).At 72 hours after surgery,the observation group had lower levels of C-reactive protein[(12.38±4.31)mg/L versus(24.87±4.68)mg/L],tumor necrosis factor-α[(9.64±1.55)μg/L versus(15.65±1.98)μg/L],and interleukin-6[(14.33±3.21)ng/mL versus(30.16±2.98)ng/mL]compared with the control group(P＜0.05).The observation group had shorter time to anal exhaust[(2.61±0.87)d versus(3.98±1.45)d],getting out of bed[(2.91±0.98)d versus(4.78±1.46)d],intake a liquid diet[(3.44±1.45)d versus(4.87±1.64)d],and first defecation[(1.66±0.18)d versus(2.93±0.78)d]compared with the control group(P＜0.05).There were no statistically significant differ-ences in the incidence of anastomotic leakage,incision infection,intra-abdominal hemorrhage,anastomotic bleeding,and reflux gastritis between the two groups(P＞0.05).Conclusion Small-incision laparoscopic radical gastrectomy can effectively reduce postoperative stress and inflammatory responses,promote postoperative recovery,and has good short-term efficacy and safety.

Objective To summarize the changing prevalence of carbapenem resistance in Enterobacterales based on the data of CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021 for improving antimicrobial treatment in clinical practice.Methods Antimicrobial susceptibility testing was performed using a commercial automated susceptibility testing system according to the unified CHINET protocol.The results were interpreted according to the breakpoints of the Clinical & Laboratory Standards Institute(CLSI)M100 31st ed in 2021.Results Over the seven-year period(2015-2021),the overall prevalence of carbapenem-resistant Enterobacterales(CRE)was 9.43％(62 342/661 235).The prevalence of CRE strains in Klebsiella pneumoniae,Citrobacter freundii,and Enterobacter cloacae was 22.38％,9.73％,and 8.47％,respectively.The prevalence of CRE strains in Escherichia coli was 1.99％.A few CRE strains were also identified in Salmonella and Shigella.The CRE strains were mainly isolated from respiratory specimens(44.23±2.80)％,followed by blood(20.88±3.40)％and urine(18.40±3.45)％.Intensive care units(ICUs)were the major source of the CRE strains(27.43±5.20)％.CRE strains were resistant to all the β-lactam antibiotics tested and most non-β-lactam antimicrobial agents.The CRE strains were relatively susceptible to tigecycline and polymyxins with low resistance rates.Conclusions The prevalence of CRE strains was increasing from 2015 to 2021.CRE strains were highly resistant to most of the antibacterial drugs used in clinical practice.Clinicians should prescribe antimicrobial agents rationally.Hospitals should strengthen antibiotic stewardship in key clinical settings such as ICUs,and take effective infection control measures to curb CRE outbreak and epidemic in hospitals.

Objective To characterize the changing species distribution and antibiotic resistance profiles of respiratory isolates in hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021.Methods Commercial automated antimicrobial susceptibility testing systems and disk diffusion method were used to test the susceptibility of respiratory bacterial isolates to antimicrobial agents following the standardized technical protocol established by the CHINET program.Results A total of 589 746 respiratory isolates were collected from 2015 to 2021.Overall,82.6％of the isolates were Gram-negative bacteria and 17.4％were Gram-positive bacteria.The bacterial isolates from outpatients and inpatients accounted for(6.0±0.9)％and(94.0±0.1)％,respectively.The top microorganisms were Klebsiella spp.,Acinetobacter spp.,Pseudomonas aeruginosa,Staphylococcus aureus,Haemophilus spp.,Stenotrophomonas maltophilia,Escherichia coli,and Streptococcus pneumoniae.Each microorganism was isolated from significantly more males than from females(P＜0.05).The overall prevalence of methicillin-resistant S.aureus(MRSA)was 39.9％.The prevalence of penicillin-resistant S.pneumoniae was 1.4％.The prevalence of extended-spectrum β-lactamase(ESBL)-producing E.coli and K.pneumoniae was 67.8％and 41.3％,respectively.The overall prevalence of carbapenem-resistant E.coli,K.pneumoniae,Enterobacter cloacae,Pseudomonas aeruginosa,and Acinetobacter baumannii was 3.7％,20.8％,9.4％,29.8％,and 73.3％,respectively.The prevalence of β-lactamase was 96.1％in Moraxella catarrhalis and 60.0％in Haemophilus influenzae.The H.influenzae isolates from children(＜18 years)showed significantly higher resistance rates to β-lactam antibiotics than the isolates from adults(P＜0.05).Conclusions Gram-negative bacteria are still predominant in respiratory isolates associated with serious antibiotic resistance.Antimicrobial resistance surveillance should be strengthened in clinical practice to support accurate etiological diagnosis and appropriate antimicrobial therapy based on antimicrobial susceptibility testing results.

With the rapid development of competitive sports, the incidence of anterior cruciate ligament (ACL) injury is on the rise. Such injuries may shorten athletes′ career and lead to other long-term adverse consequences. Although athletes generally recover well after ACL reconstruction, many still struggle to return to their pre-injury performance levels. Advances in the understanding of ACL anatomy and injury mechanisms, along with the evolution of surgical techniques and rehabilitation methods, have provided more individualized and tailored options for athletes following ACL injuries. However, there is currently no consensus in China regarding surgical and rehabilitation strategies for competitive athletes aiming to return to sports after ACL injuries. To this end, the Sports Medicine Committee of the Chinese Research Hospital Association and the Editorial Board of the Chinese Journal of Trauma jointly formulated the Expert consensus on surgical treatment and rehabilitation for competitive sports athletes returning to sports after anterior cruciate ligament injury ( version 2025), and presented 14 recommendations covering surgical indications, preoperative rehabilitation, surgical timing, surgical strategies and postoperative rehabilitation strategies, aiming to improve the surgical treatment and rehabilitation system for ACL injuries in competitive athletes and facilitate their return to high-level sports performance after injury.

BACKGROUND:At present,a large number of studies have shown that mesenchymal stem cells can be combined with different strategies to more effectively improve liver fibrosis and inhibit its progression to end-stage liver disease.OBJECTIVE:To explore the mechanism of mesenchymal stem cells combined with different strategies to improve liver fibrosis compared with mesenchymal stem cells alone.METHODS:The first author used computers to search CNKI,WanFang,VIP,PubMed,Web of Science,and Nature databases involving mesenchymal stem cells combined with different strategies to improve liver fibrosis.The search terms were"mesenchymal stem cells,liver fibrosis,combination therapy,liver stellate cells"in Chinese,and"mesenchymal stem/stromal cells,liver/hepatic fibrosis/cirrhosis,combination therapy,hepatic stellate cells/HSCs"in English.By quickly browsing the title and abstract of the article,excluding the articles that are not closely related to the topic,104 articles were finally selected for review analysis.RESULTS AND CONCLUSION:Mesenchymal stem cells improve liver fibrosis by differentiating into hepato-like cells,inhibiting hepatic stellate cell activation,immune regulation,and other mechanisms.However,the low rate of liver colonization,low survival rate,and short action time of mesenchymal stem cells after transplantation limit their clinical application.Mesenchymal stem cells can improve liver fibrosis through a combination of drugs,gene modification,cytokines and other strategies,and the efficacy is better than that of mesenchymal stem cells alone.Moreover,the combination of mesenchymal stem cells and different strategies can effectively improve liver fibrosis by promoting mesenchymal stem cell homing,inhibiting hepatic stellate cell activation,regulating the microenvironment,and regulating signaling pathways.Mesenchymal stem cells can also show better liver-derived differentiation,homing and survival functions in reducing liver fibrosis through pretreatment,miRNA regulation and combination with other cells.The combination of mesenchymal stem cells and different strategies cannot avoid the potential risks of mesenchymal stem cells alone in the treatment of liver fibrosis,and the safety of these strategies(drugs,gene modifications,cytokines,etc.)is also worth considering.In addition,the number and route of mesenchymal stem cell transplantation need to be further studied.

Objective To analyze the current status and influencing factors of occupational stress, job burnout and sleep quality among workers in the secondary industry in Jinshan District, Shanghai City. Methods A total of 1 418 workers from six key industries in Jinshan District, Shanghai City were selected as the study subjects by the stratified cluster sampling method. The Occupational Stress Core Scale, Maslash Burnout Inventory General Survey and Pittsburgh Sleep Quality Index were used to investigate occupational stress, job burnout and sleep quality of the workers. Results The detection rates of occupational stress, job burnout and sleep disturbance among the study subjects were 33.6%, 65.4% and 23.3%, respectively. Multivariate logistic regression analysis showed that the workers with a monthly income <5 000 yuan had a higher risk of occupational stress than those with a monthly income ≥5 000 yuan (P<0.01). The workers with ≥5.0 years of service had a higher risk than those with <1.0 year (P<0.05). Lack of physical exercise, employment in medium- and large-sized enterprises, and shift work were risk factors of occupational stress in the workers (all P<0.01). The workers aged 18-<30 years had a higher risk of job burnout than those aged 45-<60 years (P<0.05). The workers monthly income <5 000 yuan was associated with a higher risk of job burnout than those with ≥9 000 yuan (P<0.05). The workers with 1.0-<10.0 years or ≥15.0 years of service had higher job burnout risks than those with <1.0 year (all P<0.05). Being unmarried, lack of physical exercise, and employment in medium- and large-sized enterprises were risk factor of job burnout in the workers (all P<0.05). The workers with an educational level of high school or above had a higher risk of sleep disturbance than those with junior school or below (P<0.05). The workers who work >56 hours per week had a higher risk than those working ≤40 hours per week (P<0.01). Conclusion There is a high detection rate of occupational stress, job burnout, and sleep disturbance in the secondary industry workers in Jinshan District, Shanghai City. Special attention should be given to workers with low income, lack of physical exercise, employment in medium- and large-sized enterprises, shift work, long service duration, and long weekly working hours to protect their physical and mental health.

Objective:To study the clinical outcome of palmar approach to on-top plasty in the treatment of unequal radial polydactyly.Methods:A retrospective analysis was conducted on patients with unequal radial polydactyly in which neither thumb duplicates possess both well-developed proximal and distal components. All the patients underwent palmar approach to on-top plasty between September 2013 and September 2023 at Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine and Shanxi Children’s Hospital. At the final follow-up, the active flexion-extension range of motion of the first metacarpophalangeal and interphalangeal joints was measured, along with grip and pinch strength. The Vancouver scar scale was used to assess scar formation (total score 0-15, with higher scores indicating more severe scarring). Additionally, the pediatric quality of life inventory (total score 0-100, with higher scores indicating better quality of life, evaluated by the children themselves or by their parents/guardians on their behalf) and the satisfaction questionnaire (total score ranges from 1 to 5, with higher scores indicating greater satisfaction) were used to assess the patients’ daily quality of life and parents’satisfaction.Results:A total of 28 pediatric patients were included, comprising 15 males and 13 females, with an average age of 15 months (range: 10-36 months). All cases presented unilateral radial polydactyly, including 18 right hands and 10 left hands. The mean follow-up duration was 4.2 years (range: 1.5-6.0 years). All incisions healed in one stage, and the skin flaps survived well without infection. Joint stability was maintained without any deviation. All reconstructed thumbs demonstrated satisfactory aesthetic appearance. The reconstructed thumbs demonstrated mean active flexion-extension arcs of 76° (range 61°-85°) at the first metacarpophalangeal joint, and 42° (range 21°-65°) at the interphalangeal joint. The grip strength of the hand was 82% (63%-90%) of the normal opposite hand. The key, tip and tripod pinch strength were 76%(66%-83%), 77%(66%-85%) and 79%(68%-87%) of the normal opposite hand, respectively. No scar hyperplasia was found in all cases, and the Vancouver scar scale score was 1.2(0-3) points. The self-assessment score of the children on the pediatric quality of life inventory was 86 points, and the score of the parents/guardians was 91 points. In terms of family satisfaction, 23 patients scored 5, and 5 patients scored 4.Conclusion:The use of palmar approach to on-top plasty to treat unequal radial polydactyly can achieve functional and aesthetic results.

Host-derived small RNAs are emerging as critical regulators in the dynamic interactions between host tissues and the microbiome, with implications for microbial pathogenesis and host defense. Among these, transfer RNA-derived small RNAs (tsRNAs) have garnered attention for their roles in modulating microbial behavior. However, the bacterial factors mediating tsRNA interaction and functionality remain poorly understood. In this study, using RNA affinity pull-down assay in combination with mass spectrometry, we identified a putative membrane-bound protein, annotated as P-type ATPase transporter (PtaT) in Fusobacterium nucleatum (Fn), which binds Fn-targeting tsRNAs in a sequence-specific manner. Through targeted mutagenesis and phenotypic characterization, we showed that in both the Fn type strain and a clinical tumor isolate, deletion of ptaT led to reduced tsRNA intake and enhanced resistance to tsRNA-induced growth inhibition. Global RNA sequencing and label-free Raman spectroscopy revealed the phenotypic differences between Fn wild type and PtaT-deficient mutant, highlighting the functional significance of PtaT in purine and pyrimidine metabolism. Furthermore, AlphaFold 3 prediction provides evidence supporting the specific binding between PtaT and Fn-targeting tsRNA. By uncovering the first RNA-binding protein in Fn implicated in growth modulation through interactions with host-derived small RNAs (sRNAs), our study offers new insights into sRNA-mediated host-pathogen interplay within the context of microbiome-host interactions.
Fusobacterium nucleatum/growth & development* ; RNA-Binding Proteins/genetics* ; Bacterial Proteins/genetics* ; RNA, Bacterial/metabolism* ; Humans ; RNA, Transfer/metabolism*