Objective Recombinase-aided polymerase chain reaction(RAP)is a sensitive,single-tube,two-stage nucleic acid amplification method.This study aimed to develop an assay that can be used for the early diagnosis of three types of bacteremia caused by Staphylococcus aureus(SA),Pseudomonas aeruginosa(PA),and Acinetobacter baumannii(AB)in the bloodstream based on recombinant human mannan-binding lectin protein(M1 protein)-conjugated magnetic bead(M1 bead)enrichment of pathogens combined with RAP. Methods Recombinant plasmids were used to evaluate the assay sensitivity.Common blood influenza bacteria were used for the specific detection.Simulated and clinical plasma samples were enriched with M1 beads and then subjected to multiple recombinase-aided PCR(M-RAP)and quantitative PCR(qPCR)assays.Kappa analysis was used to evaluate the consistency between the two assays. Results The M-RAP method had sensitivity rates of 1,10,and 1 copies/μL for the detection of SA,PA,and AB plasmids,respectively,without cross-reaction to other bacterial species.The M-RAP assay obtained results for<10 CFU/mL pathogens in the blood within 4 h,with higher sensitivity than qPCR.M-RAP and qPCR for SA,PA,and AB yielded Kappa values of 0.839,0.815,and 0.856,respectively(P<0.05). Conclusion An M-RAP assay for SA,PA,and AB in blood samples utilizing M1 bead enrichment has been developed and can be potentially used for the early detection of bacteremia.

Liver cancer has the fifth morbidity and the third mortality among maligant tumors in the worldwide,and hepatocellular carci-noma(HCC)accounts for 90% of primary liver cancer.Aerobic glycolysis,as a hallmark of hepatoma cell energy metabolism,is firstly found in HCC and responsible for the regulation of cell proliferation,immune evasion,invasion,metastasis,and drug resistance.There are three rate-limiting enzymes in the glycolysis,including phosphofructokinase 1(PFK1),pyruvate kinases type M2(PKM2)and hexokinase 2(HK2),which play an important role in the regulation of aerobic glycolysis and can be regulated by many mechanisms.Because of the impor-tance of aerobic glycolysis in the occurrence and development of HCC,targeting and blocking key factors in its pathway may be a potential therapeutic treatment for HCC.With the development of key enzymes activations/inhibitors of aerobic glycolysis,the prospect of targeted therapy should be expected.Therefore,this article reviews the research progress of aerobic glycolysis in liver cancer,and summarize the role of aerobic glycolysis in the development and treatment of liver cancer,in order to provide clinical reference.

Humans ; Female ; Tibet ; Kashin-Beck Disease/epidemiology* ; China ; Metabolomics

OBJECTIVE: To examine the anti-inflammatory effect of grape seed extract (GSE) in animal and cellular models and explore its mechanism of action. METHODS: This study determined the inhibitory effect of GSE on macrophage inflammation and Th1 and Th17 polarization in vitro. Based on the in vitro results, the effects and mechanisms of GSE on multiple sclerosis (MS)-experimental autoimmune encephalomyelitis (EAE) mice model were further explored. The C57BL/6 mice were intragastrically administered with 50 mg/kg of GSE once a day from the 3rd day to the 27th day after immunization. The activation of microglia, the polarization of Th1 and Th17 and the inflammatory factors such as tumor necrosis factor- α (TNF- α), interleukin-1 β (IL-1 β), IL-6, IL-12, IL-17 and interferon-γ (IFN-γ) secreted by them were detected in vitro and in vivo by flow cytometry, enzyme linked immunosorbent assay (ELISA), immunofluorescence staining and Western blot, respectively. RESULTS: GSE reduced the secretion of TNF-α, IL-1 β and IL-6 in bone marrow-derived macrophages stimulated by lipopolysaccharide (P<0.01), inhibited the secretion of TNF-α, IL-1 β, IL-6, IL-12, IL-17 and IFN-γ in spleen cells of EAE mice immunized for 9 days (P<0.05 or P<0.01), and reduced the differentiation of Th1 and Th17 mediated by CD3 and CD28 factors (P<0.01). GSE significantly improved the clinical symptoms of EAE mice, and inhibited spinal cord demyelination and inflammatory cell infiltration. Peripherally, GSE downregulated the expression of toll-like-receptor 4 (TLR4) and Rho-associated kinase (ROCKII, P<0.05 or P<0.01), and inhibited the secretion of inflammatory factors (P<0.01 or P<0.05). In the central nervous system, GSE inhibited the infiltration of CD45⁺CD11b⁺ and CD45⁺CD4⁺ cells, and weakened the differentiation of Th1 and Th17 (P<0.05). Moreover, it reduced the secretion of inflammatory factors (P<0.01), and prevented the activation of microglia (P<0.05). CONCLUSION GSE had a beneficial effect on the pathogenesis and progression of EAE by inhibiting inflammatory response as a potential drug and strategy for the treatment of MS.
Mice ; Animals ; Encephalomyelitis, Autoimmune, Experimental/pathology* ; Grape Seed Extract/therapeutic use* ; Interleukin-17 ; Interleukin-1beta ; Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-6/metabolism* ; Th1 Cells ; Mice, Inbred C57BL ; Interferon-gamma/therapeutic use* ; Th17 Cells/metabolism* ; Interleukin-12/therapeutic use* ; Cytokines/metabolism*

Objective: To explore the molecular features of chronic myelomonocytic leukemia (CMML) . Methods: According to 2022 World Health Organization (WHO 2022) classification, 113 CMML patients and 840 myelodysplastic syndrome (MDS) patients from March 2016 to October 2021 were reclassified, and the clinical and molecular features of CMML patients were analyzed. Results: Among 113 CMML patients, 23 (20.4%) were re-diagnosed as acute myeloid leukemia (AML), including 18 AML with NPM1 mutation, 3 AML with KMT2A rearrangement, and 2 AML with MECOM rearrangement. The remaining 90 patients met the WHO 2022 CMML criteria. In addition, 19 of 840 (2.3%) MDS patients met the WHO 2022 CMML criteria. At least one gene mutation was detected in 99% of CMML patients, and the median number of mutations was 4. The genes with mutation frequency ≥ 10% were: ASXL1 (48%), NRAS (34%), RUNX1 (33%), TET2 (28%), U2AF1 (23%), SRSF2 (21.1%), SETBP1 (20%), KRAS (17%), CBL (15.6%) and DNMT3A (11%). Paired analysis showed that SRSF2 was frequently co-mutated with ASXL1 (OR=4.129, 95% CI 1.481-11.510, Q=0.007) and TET2 (OR=5.276, 95% CI 1.979-14.065, Q=0.001). SRSF2 and TET2 frequently occurred in elderly (≥60 years) patients with myeloproliferative CMML (MP-CMML). U2AF1 mutations were often mutually exclusive with TET2 (OR=0.174, 95% CI 0.038-0.791, Q=0.024), and were common in younger (<60 years) patients with myelodysplastic CMML (MD-CMML). Compared with patients with absolute monocyte count (AMoC) ≥1×10(9)/L and <1×10(9)/L, the former had a higher median age of onset (60 years old vs 47 years old, P<0.001), white blood cell count (15.9×10(9)/L vs 4.4×10(9)/L, P<0.001), proportion of monocytes (21.5% vs 15%, P=0.001), and hemoglobin level (86 g/L vs 74 g/L, P=0.014). TET2 mutations (P=0.021) and SRSF2 mutations (P=0.011) were more common in patients with AMoC≥1×10(9)/L, whereas U2AF1 mutations (P<0.001) were more common in patients with AMoC<1×10(9)/L. There was no significant difference in the frequency of other gene mutations between the two groups. Conclusion: According to WHO 2022 classification, nearly 20% of CMML patients had AMoC<1×10(9)/L at the time of diagnosis, and MD-CMML and MP-CMML had different molecular features.
Humans ; Aged ; Middle Aged ; Leukemia, Myelomonocytic, Chronic/genetics* ; Prognosis ; Splicing Factor U2AF/genetics* ; Mutation ; Myelodysplastic Syndromes/genetics* ; Leukemia, Myeloid, Acute/genetics*

Objective: To methodically assess the clinical effectiveness and safety of robot-assisted total rectal mesenteric resection (RTME), laparoscopic-assisted total rectal mesenteric resection (laTME), and transanal total rectal mesenteric resection (taTME). Methods: A computer search was conducted on PubMed, Embase, Cochrane Library, and Ovid databases to identify English-language reports published between January 2017 and January 2022 that compared the clinical efficacy of the three surgical procedures of RTME, laTME, and taTME. The quality of the studies was evaluated using the NOS and JADAD scales for retrospective cohort studies and randomized controlled trials, respectively. Direct meta-analysis and reticulated meta-analysis were performed using Review Manager software and R software, respectively. Results: Twenty-nine publications comprising 8,339 patients with rectal cancer were ultimately included. The direct meta-analysis indicated that the length of hospital stay was longer after RTME than after taTME, whereas according to the reticulated meta-analysis the length of hospital stay was shorter after taTME than after laTME (MD=-0.86, 95%CI: -1.70 to -0.096, P=0.036). Moreover, the incidence of anastomotic leak was lower after taTME than after RTME (OR=0.60, 95%CI: 0.39 to 0.91, P=0.018). The incidence of intestinal obstruction was also lower after taTME than after RTME (OR=0.55, 95%CI: 0.31 to 0.94, P=0.037). All of these differences were statistically significant (all P<0.05). There were no statistically significant differences between the three surgical procedures regarding the number of lymph nodes cleared, length of the inferior rectal margin, or rate of positive circumferential margins (all P>0.05). An inconsistency test using nodal analysis revealed no statistically significant differences between the results of direct and indirect comparisons of the six outcome indicators (all P>0.05). Furthermore, we detected no significant overall inconsistency between direct and indirect evidence. Conclusion: taTME has advantages over RTME and laTME, in terms of radical and surgical short-term outcomes in patients with rectal cancer.
Humans ; Robotics ; Robotic Surgical Procedures/adverse effects* ; Network Meta-Analysis ; Retrospective Studies ; Postoperative Complications/etiology* ; Transanal Endoscopic Surgery/methods* ; Rectum/surgery* ; Rectal Neoplasms/pathology* ; Laparoscopy/methods* ; Treatment Outcome

Objective:To investigate the effect of fat mass index (FMI) on early recovery after total knee arthroplasty (TKA).Methods:Patients who underwent primary unilateral TKA in Xi'an Honghui Hospital from July 2020 to July 2021 were retrospectively analyzed. The preoperative body composition was measured by dual energy X-ray absorptiometry and the FMI was calculated. Patients were divided into normal group (male: 3.0-6.0 kg/m 2; female: 5.0-9.0 kg/m 2), overweight group (male: 6.1-9.0 kg/m 2; female: 9.1-13.0 kg/m 2), and obese group (male: >9 kg/m 2; female: >13 kg/m 2) according to level of FMI, and the operation time, blood loss, and incidence of postoperative complications were collected. Multifactorial analysis of the effect of FMI on early recovery after TKA was performed using a generalized linear model. Draw the receiver operating characteristics (ROC) curve of BMI and FMI on the predicted effect of postoperative Western Ontario and McMaster Universities (WOMAC) osteoarthritis index scores and Knee Society Score (KSS) to compare the effect of FMI with BMI on early recovery after TKA. Results:A total of 100 patients were included in the study, 24 males and 76 females, aged 65.0±8.2 years (range, 42-81 years). There were 15 cases in normal group, 55 cases in overweight group and 30 cases in obese group. All patients successfully completed the operation and were followed up for 3.15±0.72 months (range, 2.8-3.2 months). The WOMAC scores of the obese group at 2 weeks, 1 and 2 months postoperative were 34.57±3.68, 22.03±2.79, and 15.77±2.96, which were greater than those of the normal group (28.73 ±2.58, 19.07±2.71, 12.27±3.10), as well as the overweight group (30.05±4.09, 19.33±2.42, 14.84±2.42), with statistically significant differences ( P<0.05). The KSS scores of the obese group at postoperative 1 and 2 months were 68.83±5.52 and 81.17±4.49, which were lower than those of the normal group (77.33±5.63, 87.33±4.17), as well as the overweight group (72.64±5.43, 83.73 ±5.02), with statistically significant differences ( P<0.05). The WOMAC score, KSS score, and postoperative complications at 2 months postoperatively were selected as outcome indicators to plot the ROC curve, and the ROC curve for the WOMAC score at 2 months postoperatively showed an area under the curve corresponding to FMI of 0.744 (95% CI: 0.54, 0.82), which was greater than that of BMI [0.624 (95% CI: 0.51, 0.74)], and the difference was statistically significant ( Z=2.19, P=0.021). The ROC curve for the KSS score at 2 months postoperatively showed an area under the curve corresponding to FMI of 0.718 (95% CI: 0.62, 0.82), which was greater than that of BMI [0.612 (95% CI: 0.52, 0.74)], with a statistically significant difference ( Z=2.58, P=0.016). The ROC curve for postoperative complications showed an area under the curve of 0.639 (95% CI: 0.41, 0.88) for FMI and 0.605 (95% CI: 0.37, 0.84) for BMI, with no statistically significant difference ( Z=0.48, P=0.632). Conclusion:The greater the FMI the poorer the early functional recovery after initial TKA, and FMI is more valuable than BMI in predicting the early functional recovery.

Objective To analyze the plantar pressure distribution of knee osteoarthritis ( KOA) patients after medial opening wedge high tibial osteotomy ( MOWHTO), so as to provide biomechanical references for the surgical treatment and rehabilitation of patients. Methods A total of 31 patients with medial single compartmental KOA after unilateral MOWHTO treatment were selected as the experimental group, and 35 healthy subjects at same age were selected as the control group. The Pedomedic 40 􀅺 pressure measuring system was used to test dynamic plantar pressure. By comparing the maximum pressure ( pmax ), force-time integral ( FTI) and contact area (CA) of different plantar zones between the experimental group (operative side and unoperated side) and the control group during walking, the changes of plantar pressure in patients with medial single compartmental KOA after MOWHTO were evaluated. Results Compared with the unoperated side and the control group, the CA and FTI of the 1st metatarsal head (MH1) were higher (P<0. 05), the CA of the 4th metatarsal head (MH4)was smaller (P<0. 001), the pmax and FTI of the 5th metatarsal head (MH5) were smaller (P<0. 05), the CA of the lateral middle foot (MF-L) was smaller (P<0. 001), and the CA of the medial rear foot (RF-M) was larger (P<0. 05). Compared with the control group, the pmax of MH1 and MH2 was smaller (P<0. 05), the CA and FTI of MH5 were larger (P<0. 05), the pmax of MF-L was larger (P<0. 001), and the FTI of lateral rear foot (RF-L) was larger (P<0. 05). Conclusions Compared with healthy people, patients with medial single compartmental KOA have abnormal plantar pressure residual after MOWHTO. In clinical practice, targeted intensive rehabilitation therapy is necessary to restore the normal plantar distributions of patients.

Primary ciliary dyskinesia (PCD) is a rare hereditary orphan condition that results in variable phenotypes, including infertility. About 50 gene variants are reported in the scientific literature to cause PCD, and among them, dynein axonemal assembly factor 4 ( DNAAF4 ) has been recently reported. DNAAF4 has been implicated in the preassembly of a multiunit dynein protein essential for the normal function of locomotory cilia as well as flagella. In the current study, a single patient belonging to a Chinese family was recruited, having been diagnosed with PCD and asthenoteratozoospermia. The affected individual was a 32-year-old male from a nonconsanguineous family. He also had abnormal spine structure and spinal cord bends at angles diagnosed with scoliosis. Medical reports, laboratory results, and imaging data were investigated. Whole-exome sequencing, Sanger sequencing, immunofluorescence analysis, hematoxylin-eosin staining, and in silico functional analysis, including protein modeling and docking studies, were used. The results identified DNAAF4 disease-related variants and confirmed their pathogenicity. Genetic analysis through whole-exome sequencing identified two pathogenic biallelic variants in the affected individual. The identified variants were a hemizygous splice site c.784-1G>A and heterozygous 20.1 Kb deletion at the DNAAF4 locus, resulting in a truncated and functionless DNAAF4 protein. Immunofluorescence analysis indicated that the inner dynein arm was not present in the sperm flagellum, and sperm morphological analysis revealed small sperm with twisted and curved flagella or lacking flagella. The current study found novel biallelic variants causing PCD and asthenoteratozoospermia, extending the range of DNAAF4 pathogenic variants in PCD and associated with the etiology of asthenoteratozoospermia. These findings will improve our understanding of the etiology of PCD.
Adult ; Humans ; Male ; Asthenozoospermia/genetics* ; Dyneins/genetics* ; East Asian People ; Kartagener Syndrome/genetics* ; Mutation ; Proteins/genetics* ; Semen/metabolism*

Objective: To investigate the efficacy and safety of pedunculated rectus abdominis combined with bilateral ureteral extravestheter drainage in the treatment of refractory bladder-vaginal stump fistula. Methods: The clinical data of 8 cases of the refractory bladder-vaginal stump fistula were admitted to the Second Hospital of Hebei Medical University and Henan Cancer Hospital and underwent the clinical treatment of bladder-vaginal stump from December 2019 to December 2022 were collected. The reason of refractory bladder-vaginal stump fistula was analyzed, the operation manner of pedunculated rectus abdominis combined with peduncle and bilateral ureter for the treatment of bladder-vaginal stump through extrabladder drainage was explored. The operation time, bleeding volume and clinical effect were record. Results: The median operation time of 8 patients was 150 minutes(120~180 min), and the median blood loss was 400 ml(200~600 ml). During the perioperative period, there were 2 cases of incision infection, delayed healing by debridement and dressing, 2 cases of incision rupture and suture wound healing after reoperation, and 2 cases of urinary tract infection were cured by anti-infection. When followed up for 6 months, 8 cases of vesicovaginal stump fistula were cured. Conclusion: Bilateral ureteral external drainage of the rectus abdominis muscle, has a practical effect in the treatment of refractory bladder-vaginal stump fistula, which can be one of the clinical repairing treatment.
Female ; Humans ; Urinary Bladder/surgery* ; Ureter/surgery* ; Rectus Abdominis ; Drainage ; Fistula