Efficient site-specific integration of large genes in mammalian cells via continuously evolved recombinases and prime editing.

Methods for the targeted integration of genes in mammalian genomes suffer from low programmability, low efficiencies or low specificities. Here we show that phage-assisted continuous evolution enhances prime-editing-assisted site-specific integrase gene editing (PASSIGE), which couples the programmability of prime editing with the ability of recombinases to precisely integrate large DNA cargoes exceeding 10 kilobases. Evolved and engineered Bxb1 recombinase variants (evoBxb1 and eeBxb1) mediated up to 60% donor integration (3.2-fold that of wild-type Bxb1) in human cell lines with pre-installed recombinase landing sites. In single-transfection experiments at safe-harbour and therapeutically relevant sites, PASSIGE with eeBxb1 led to an average targeted-gene-integration efficiencies of 23% (4.2-fold that of wild-type Bxb1). Notably, integration efficiencies exceeded 30% at multiple sites in primary human fibroblasts. PASSIGE with evoBxb1 or eeBxb1 outperformed PASTE (for 'programmable addition via site-specific targeting elements', a method that uses prime editors fused to recombinases) on average by 9.1-fold and 16-fold, respectively. PASSIGE with continuously evolved recombinases is an unusually efficient method for the targeted integration of genes in mammalian cells.

[Analysis of a family with 11ß-hydroxylase deficiency due to a mutation in the CYP11B1 gene].

This study reports a family of patients with 11ß-hydroxylase deficiency (11ß-OHD) caused by a novel mutation in the CYP11B1 gene, and analyzes its clinical and genetic characteristics. The clinical data of a patient with intractable hypertension at Air Force Medical Center on May 16, 2014 were retrospectively analyzed. The patient was clinically diagnosed with congenital adrenal cortical hyperplasia. The clinical data of the patient were further collected and the peripheral blood samples of the patient, his parents and his sister were collected for CYP11B1(NM_000497) gene sequencing, suggesting that the patient had compound heterozygous mutations in exon 1:c.199delG, p.Glu67Lysfs*9 and exon 5:c.905_907 delATGinsTT, p.Asp302Valfs*23, both of which were pathogenic variants. The patient's father and sister carried heterozygous mutations in exon 1:c.199delG, p.Glu67Lysfs*9, and the mother carried heterozygous mutations in exon 5:c.905_907delATGinsTT, p.Asp302Valfs*23. This study is the first to report a new compound heterozygous mutation in exon 1:c.199delG and exon 5 c.905_907 delATGinsTT of CYP11B1 gene, enriching the database of 11ß-OHD mutations and providing information to further understand the genetic mechanism of the disease.

Association of noise exposure with lipid metabolism among Chinese adults: mediation role of obesity indices.

PURPOSE: Noise exposure in the workplace has been linked to a number of health consequences. Our objectives were to explore the relationship between occupational noise and lipid metabolism and evaluate the possible mediating effect of obesity indices in those relationships with a cross-sectional study design. METHODS: Cumulative noise exposure (CNE) was used to measure the level of noise exposure. Logistic regression models or generalized linear models were employed to evaluate the association of occupational noise and obesity with lipid metabolism markers. Cross-lagged analysis was conducted to explore temporal associations of obesity with lipid metabolism. RESULTS: A total of 854 participants were included, with each one-unit increase in CNE, the values of total cholesterol/high-density lipoprotein cholesterol and low-density lipoprotein cholesterol/high-density lipoprotein cholesterol increased by 0.013 (95% confidence interval: 0.006, 0.020) and 0.009 (0.004, 0.014), as well as the prevalence of dyslipidemia increased by 1.030 (1.013, 1.048). Occupational noise and lipid metabolism markers were all positively associated with body mass index (BMI), waist circumference (WC), a Body Shape Index (ABSI) and a Body Shape Index and Body Roundness Index (BRI) (all P < 0.05). Moreover, BMI, WC, ABSI and BRI could mediate the associations of occupational noise with lipid metabolism; the proportions ranged from 21.51 to 24.45%, 23.84 to 30.14%, 4.86 to 5.94% and 25.59 to 28.23%, respectively (all P < 0.05). CONCLUSIONS: Our study demonstrates a positive association between occupational noise and abnormal lipid metabolism, and obesity may partly mediate the association. Our findings reinforce the need to take practical steps to reduce or even eliminate the health risks associated with occupational noise.

Clinical efficacy and safety evaluation of Dermatophagoides farinae drops in the treatment of allergic rhinitis with epistaxis.

Summary: Background. Epistaxis is frequently observed in allergic rhinitis (AR) patients. However, few studies focus on the outcome of epistaxis with treatment of AR patients. This study aimed to retrospectively analyze the efficacy and safety of AR patients with epistaxis treated with sublingual immunotherapy (SLIT). Methods. A total of 74 patients aged 4-60 years with house dust mite (HDM)-induced AR accompanied by epistaxis and who completed 1 year of SLIT treatment with standard Dermatophagoides farinae (D. farinae) drops were enrolled in this study. The symptom scores, total medication scores (TMS), combined symptom and medication score (CSMS), visual analog scales (VAS), and bleeding score (BS) were assessed, as well as the nasal endoscopic examinations were performed to observe nasal signs. Results. The levels of symptom scores, TMS, CSMS, VAS, and BS at 0.5 year and 1 year of SLIT treatment were significantly lower than those at the baseline (all p less than 0.01). Also, statistical differences were seen in CSMS (p less than 0.05) and VAS (p less than 0.01) between 0.5 year and 1 year. As expected, BS was positively correlated with CSMS (r = 0.617, 95% CI 0.517-0.699) and VAS (r = 0.777, 95% CI 0.719-0.822) at all three time points. Conclusions. SLIT with D. farinae drops was effective and safe for AR patients with epistaxis, resulting in improving the symptoms of rhinitis while relieving the symptoms of epistaxis.

Airborne antibiotic and metal resistance genes - A neglected potential risk at e-waste recycling facilities.

Heavy metal-rich environments can promote the selection of metal-resistance genes (MRGs) in bacteria, often leading to the simultaneous selection of antibiotic-resistance genes (ARGs) through a process known as co-selection. To comprehensively evaluate the biological pollutants at electronic-waste (e-waste) recycling facilities, air, soil, and river samples were collected at four distinct Swiss e-waste recycling facilities and analyzed for ARGs, MRGs, mobile genetic elements (MGEs), endotoxins, and bacterial species, with correlations drawn to heavy metal occurrence. To our knowledge, the present work marks the first attempt to quantify these bio-pollutants in the air of e-waste recycling facilities, that might pose a significant health risk to workers. Although ARG and MRG's profiles varied among the different sample types, intl1 consistently exhibited high relative abundance rates, identifying it as the predominant MGE across all sample types and facilities. These findings underscore its pivol role in driving diverse bacterial adaptations to extreme heavy metal exposure by selection and dissemination of ARGs and MRGs. All air samples exhibited consistent profiles of ARGs and MRGs, with blaTEM emerging as the predominant ARG, alongside pbrT and nccA as the most prevalent MRGs. However, one facility, engaged in batteries recycling and characterized by exceptionally high concentrations of heavy metals, showcased a more diverse resistance gene profile, suggesting that bacteria in this environment required more complex resistance mechanisms to cope with extreme metal exposure. Furthermore, this study unveiled a strong association between gram-negative bacteria and ARGs and less with MRGs. Overall, this research emphasizes the critical importance of studying biological pollutants in the air of e-waste recycling facilities to inform robust safety measures and mitigate the risk of resistance gene dissemination among workers. These findings establish a solid foundation for further investigations into the complex interplay among heavy metal exposure, bacterial adaptation, and resistance patterns in such distinctive ecosystems.

Cutaneous metastasis from gastric cancer: Manifestation, diagnosis, treatment and prognosis.

INTRODUCTION: Cutaneous metastasis from gastric cancer is very rare. The understanding of this disease is incomplete. This situation delays its diagnosis and treatment, followed by poor prognosis. Here, we first report a study based on a network to improve the diagnosis, treatment and prognosis of cutaneous metastasis from gastric cancer. METHODS: A comprehensive search of PubMed was performed. All studies on cutaneous metastasis from gastric cancer were collected. The publication date was limited from 2000 to the present, and the language was limited to English. SPSS 26.0 was employed for statistical analysis. RESULTS: Seventy-two patients were included. The average patient age was 60.0 ± 16.0 years. In total, 72.2 % of the patients were male. The most common manifestation was nodular skin lesions (45.8 %). The metastases generally presented as multiple lesions (61.1 %). The most common metastasis location was the thoracoabdominal wall (56.9 %). 64.7 % of the patients simultaneously had extracutaneous metastases. Most of the tumors were poorly differentiated carcinomas (87.5 %), and 66.1 % had signet ring cells. 40.8 % of the cutaneous metastases presented as primary manifestations. Only 9.6 % had their diagnosis as soon as the cutaneous metastasis emerged. Systemic chemotherapy (65.6 %) was the most common treatment strategy, followed by radical surgery (12.5 %). The median overall survival was only 6 months. The median overall survival of 5 patients with resected tumors was 48 months. CONCLUSION: Cutaneous metastasis from gastric cancer usually manifests as an emerged nodule or erysipelas-like skin lesion. Resection of the cutaneous lesion could be helpful for patients with local metastases.

[A family with developmental glaucoma and microcornea due to novel ADAMTS18 gene mutations].

This case report presents a family with developmental glaucoma accompanied by microcornea resulting from novel mutations in the ADAMTS18 gene. The index case involves a 5-year-old twin brother, who, during a routine examination, exhibited elevated intraocular pressure persisting for over a month. The peak intraocular pressure reached approximately 25 mmHg (1 mmHg=0.133 kPa) in both eyes, with a corneal diameter of less than 10 mm. Ocular examination revealed an enlarged cup-to-disc ratio, and optical coherence tomography (OCT) demonstrated thinning of the retinal nerve fiber layer and ganglion cell layer. Ultrasound biomicroscopy combined with gonioscopy indicated partial angle closure and abnormal anterior chamber angle development. The ocular manifestations in the twin brother were consistent with those observed in the twin sister. The clinical diagnosis was bilateral developmental glaucoma with microcornea. Genetic sequencing identified two novel compound heterozygous mutations in the ADAMTS18 gene in the twins: Mutation 1 (M1) involving the variant site 1 (c.3436C>T:p.R1146W) and Mutation 2 (M2) involving the variant site 2 (c.1454T>G:p.F485C). Ocular examinations of four additional family members were normal. Genetic testing revealed that the twins' father and sister carried M1, while the index case's mother and brother carried M2. This report underscores a unique association between ADAMTS18 gene mutations and developmental glaucoma with microcornea within a familial context, emphasizing the importance of genetic screening for early diagnosis and targeted management strategies.

[Epidemiological characteristics of a local cluster epidemic caused by the BA.2 evolutionary branch of Omicron variant].

Descriptive epidemiological methods were used to analyze the epidemiological characteristics of the local cluster of COVID-19 in the logistic park of Yuhang District in Hangzhou in March 2022. The cluster epidemic was detected by a case who actively visited the fever clinic. The epidemic lasted for 8 days, and a total of 58 cases (53 workers, 2 students, 1 farmer, 1 teacher and 1 unemployed) were found, including 40 males and 18 females. The age was (33.29±12.22) years. There cases were mainly in Yuhang District (48 cases, 82.77%) and Shangcheng District (7 cases, 12.07%) of Hangzhou. The real-time regeneration number peaked at 2.31 on March 10th and decreased to 0.37 on March 15th. The sequencing result of the indicated case was 100% homologous with the sequence uploaded from South Korea on March 4th, 2022.

Efficacy of acupuncture in the treatment of limb dyskinesia after stroke: a systematic review and meta-analysis.

OBJECTIVE: This meta-analysis was performed to investigate the effectiveness of acupuncture in post-stroke limb movement disorders. MATERIALS AND METHODS: An electronic search of databases including MEDLINE/PubMed, Web of Science, the Cochrane database, EMBASE, CBM, CNKI, WanFang, and VIP was performed to collect randomized controlled clinical studies on acupuncture administered for post-stroke dyskinesia from inception to April 2023. Data including baseline information, Fugl-Meyer Assessment (FMA) scores, and Barthel Index (BI) were included and analyzed using the meta package in R language. RESULTS: After searching and screening, 17 pieces of literature involving 1,928 participants were included, with 962 participants in the control group and 966 in the study group. Results from the included studies suggested significant benefits provided by acupuncture to improve FMA scores and BI. In specific, incorporation of acupuncture in the treatment of post-stroke limb movement disorders significantly reduced the overall FMA scores of patients by 3.45 (95% CI: 0.22, 6.69) points, the upper extremity FMA scores by 3.63 (95% CI: 0.64, 6.62) points, the lower extremity FMA scores by 3.56 (95% CI: 1.78, 5.35) points, and BI by 7.75 (95% CI: 3.35, 12.16) points. CONCLUSIONS: Acupuncture as an adjunct to the management of post-stroke limb movement disorders contributes significantly to enhancing the motor function and quality of life of patients. However, the evidence of this study is compromised by the limited quantity of the included randomized controlled trials (RCTs) and the mediocre methodological quality. Therefore, high-quality randomized controlled trials are required to validate the benefits of acupuncture on the motor function of patients with post-stroke limb movement disorders.

[The efficacy analysis of neurosurgical robot-assisted DBS in the treatment of elderly Parkinson's disease].

Objective: To investigate the surgical efficacy of neurosurgery robot deep brain stimulation(DBS) in the treatment of elderly Parkinson's disease(PD). Methods: The clinical data of elderly patients (≥75 years) with PD who underwent neurosurgical robot-assisted DBS surgery in the Department of Neurosurgery of the General Hospital of Northern Theater Command from September 2016 to September 2022 were collected retrospectively. Operation time, electrode implantation duration, postoperative pneumocephalus volume, electrode implantation accuracy, the Tao's DBS surgery scale, perioperative complications were analyzed.The unified Parkinson's disease rating scales (UPDRS), UPDRS-Ⅲ, tremor, rigidity, bradykinesia, axial, Barthel Activities of Daily Living (ADL-Barthel), Levodopa Equivalent Daily Dose (LEDD), Montreal Cognitive Assessment (MoCA), Hamilton Anxiety Scale (HAMA) and Hamilton Depression Scale (HAMD) scores and mortality were assessed respectively before operation, 6, 12 and 24 months after operation and last follow-up. Results: A total of 25 elderly patients were enrolled, including 14 males and 11 females, aged(78.3±3.2) years. Nine patients had underlying diseases. Nine patients (36%) underwent bilateral Globus Pallidus pars Interna deep brain stimulation (GPi-DBS) and 16 patients (64%) underwent bilateral subthalamic nucleus deep brain stimulation (STN-DBS).The operation time was (1.56±0.19) hours, the electrode implantation duration was (1.01±0.19) hours, the pneumocephalus volume was 9.8(4.7, 23.3) cm3, and the electrode implantation accuracy was (0.84±0.24) mm, the Tao's DBS surgery scale was (80.2±6.2).The follow-up time [M(Q1, Q3)] was 57.3(27.9, 75.7) months. No serious complications such as intracranial hemorrhage, infection or poor wound healing occurred during the perioperative period. The improvement rate of UPDRS, UPDRS-Ⅲ, rigidity, bradykinesia, and LEDD at 6 months after surgery was significantly higher than that at 24 months after surgery and at the last follow-up (all P<0.05); the improvement rate of axial symptoms, ADL-Barthel score, and MoCA score at 6 months after surgery was significantly higher than that at the last follow-up (P<0.05). HAMD and HAMA scores showed no significant improvement during follow-up after surgery (both P>0.05). At the last follow-up, 12 patients died, with death time of (35.1±20.2) months after operation, and the death age of [M(Q1, Q3)] 80(79, 83)years. Conclusions: Robot-assisted DBS surgery for elderly patients with PD is accurate and safe, and the postoperative symptoms are significantly improved, and they can benefit from neuromodulation for long term, and the risks are controllable.

[Research progress of biomaterials with ordered micro-nano structure in wound repair].

Many injury-causing factors, including burns and surgery, etc., can lead to the destruction of structure and function of skin. Suitable wound dressing or implant is the material basis to promote wound healing and regeneration. Biomaterials with micro-nano structure can affect cell behavior, promote orderly growth of cell in accordance with the structure. Their application in wound healing can promote angiogenesis, regulate immune response, and reduce scar area. In recent years, the application of biomaterials with ordered micro-nano structure in tissue engineering has attracted extensive attention. This paper introduces the structure and preparation methods of several biomaterials with ordered micro-nano structure, and focuses on how the surface microstructure of biomaterials affects the process of wound healing and its molecular mechanism, in order to find and develop medical biomaterials that are closer to the skin tissue structure for clinical wound treatment.

[Near-infrared light therapy ameliorates depression-induced intestinal dysfunction in rats possibly by activating PGC-1α/Nrf2 signaling and increasing hippocampal BDNF expression].

OBJECTIVE: To investigate the effect of near-infrared (NIR) light therapy on depression-induced intestinal dysfunction in rats and explore the possible mechanism. METHODS: Thirty-two male SD rats were randomly divided into control group, model group, low-dose NIR light group and high-dose NIR light group. All the rats except for those in the control group were subjected to chronic restrained stress (CRS) for 4 weeks, and NIR light therapy of the head was administered in the two NIR light groups. The depression- like behaviors, intestinal functions, fecal water content and number of fecal pellets of the rats were evaluated. HE staining was used for detecting histopathological changes in the hippocampus and colon, and hippocampal expressions of BDNF, Nrf2 and PGC-1α were detected with Western blotting. RESULTS: The rats in the CRS model group showed significantly increased immobility time and visceral sensitivity in the behavioral tests, decreased fecal pellets and fecal water content, and lowered expressions of BDNF, Nrf2, and PGC-1α in the hippocampus (P<0.05). Histopathological examination of the CRS rats revealed loosely arranged hippocampal pyramidal cells, obvious neuronal damages, and obvious inflammatory cell infiltration in the colon with irregularly arranged mucosal glands and a high pathological score. High-dose NIR light therapy significantly lowered the immobility time and visceral sensitivity, increased the number of fecal pellets and fecal water content (P<0.05), and enhanced hippocampal expressions of BDNF, Nrf2, and PGC-1α (P<0.05) of the depressive rats. The rats receiving high-dose NIR light therapy also exhibited close arrangement of the hippocampal pyramidal cells with significantly reduced neuronal damage and colonic inflammatory cell infiltration, neatly arranged mucosal glands, and lowered pathological score. CONCLUSION: NIR light therapy can significantly improve depression-like behavior and intestinal function in rats possibly by ameliorating oxidative stress via the PGC-1α/Nrf2 signaling pathway and increasing BDNF level in the hippocampus.

Phage-assisted evolution and protein engineering yield compact, efficient prime editors.

Prime editing enables a wide variety of precise genome edits in living cells. Here we use protein evolution and engineering to generate prime editors with reduced size and improved efficiency. Using phage-assisted evolution, we improved editing efficiencies of compact reverse transcriptases by up to 22-fold and generated prime editors that are 516-810 base pairs smaller than the current-generation editor PEmax. We discovered that different reverse transcriptases specialize in different types of edits and used this insight to generate reverse transcriptases that outperform PEmax and PEmaxΔRNaseH, the truncated editor used in dual-AAV delivery systems. Finally, we generated Cas9 domains that improve prime editing. These resulting editors (PE6a-g) enhance therapeutically relevant editing in patient-derived fibroblasts and primary human T-cells. PE6 variants also enable longer insertions to be installed in vivo following dual-AAV delivery, achieving 40% loxP insertion in the cortex of the murine brain, a 24-fold improvement compared to previous state-of-the-art prime editors.

[Posterior arthroscopic subtalar arthrodesis for symptomatic adult talocalcaneal coalition].

Objective: To investigate the functional outcomes of posterior arthroscopic subtalar arthrodesis (PASTA) for adult patients presenting with symptomatic talocalcaneal coalition. Methods: The study was a retrospective case-series research.The data of 17 adult patients (17 feet) with symptomatic talocalcaneal coalitions,treated with PASTA from March 2018 to February 2022 in Xuzhou Central Hospital were collected.This procedure involved 10 males and 7 females,aged (42.4±7.5) years(range:31 to 58 years).There were 9 cases on the right side and 7 cases on the left side.According to the Rozansky classification,there were 4 cases of type Ⅰ,7 cases of type Ⅱ, 3 cases of type Ⅲ,3 cases of type Ⅳ.The following items such as wound healing and bony union of the subtalar joint were observed.Clinical assessment was performed using pain visual analogue scale (VAS),American Orthopaedic Foot & Ankle Society (AOFAS) ankle-hindfoot scores and 36-item short form health survey (SF-36) scores.The paired t test was used for data comparison. Results: The follow-up time was (24.8±6.9) months(range:12 to 40 months).There were no complications such as wound infection,deep vein thrombosis,nonunion,or screw breakage.One patient with preoperative spasm,relieved after the second surgical procedure (peroneal brevis tendon lengthening).The union time of the subtalar joint was (8.8±2.2) weeks(range:6 to 12 weeks).At the final follow-up,the VAS decreased from (6.4±1.3) to (1.3±0.9)(t=14.114,P<0.01), the AOFAS ankle-hindfoot score increased from (49.0±8.1) to (90.0±5.1)(t=38.782,P<0.01),and the SF-36 score increased from (50.8±9.5) to (91.0±4.9)(t=20.468,P<0.01). Conclusion: PASTA for adult patients presenting with symptomatic talocalcaneal coalition offers advantages of minimal trauma,fast recovery,and few complications,which is an effective method.

Spatial and temporal distribution of endotoxins, antibiotic resistance genes and mobile genetic elements in the air of a dairy farm in Germany.

Antimicrobial resistance (AMR) is a serious issue that is continuously growing and spreading, leading to a dwindling number of effective treatments for infections that were easily treatable with antibiotics in the past. Animal farms are a major hotspot for AMR, where antimicrobials are often overused, misused, and abused, in addition to overcrowding of animals. In this study, we investigated the risk of AMR transmission from a farm to nearby residential areas by examining the overall occurrence of endotoxins, antibiotic resistance genes (ARGs), and mobile genetic elements (MGEs) in the air of a cattle farm. We assessed various factors, including the season and year, day and nighttime, and different locations within the farm building and its vicinity. The most abundant ARGs detected were tetW, aadA1, and sul2, genes that encode for resistances towards antibiotics commonly used in veterinary medicine. While there was a clear concentration gradient for endotoxin from the middle of the farm building to the outside areas, the abundance of ARGs and MGEs was relatively uniform among all locations within the farm and its vicinity. This suggests that endotoxins preferentially accumulated in the coarse particle fraction, which deposited quickly, as opposed to the ARGs and MGEs, which might concentrate in the fine particle fraction and remain longer in the aerosol phase. The occurrence of the same genes found in the air samples and in the manure indicated that ARGs and MGEs in the air mostly originated from the cows, continuously being released from the manure to the air. Although our atmospheric dispersion model indicated a relatively low risk for nearby residential areas, farm workers might be at greater risk of getting infected with resistant bacteria and experiencing overall respiratory tract issues due to continuous exposure to elevated concentrations of endotoxins, ARGs and MGEs in the air of the farm.

[Clinical analysis of nodular fasciitis in the head and neck].

Objective: To analyze the clinical features, diagnosis, treatment and prognosis of nodular fasciitis (NF) in the head and neck. Methods: Seven cases of primary NF in the head and neck admitted to Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from 1990 to 2022 were retrospectively analyzed, including 3 males and 4 females, aged from 2 to 67 years, and the location, course of disease, clinical manifestations, imaging findings, and treatment results of the disease were summarized. Results: Of the 7 patients, the primary sites were respectively nasal vestibule, paranasal sinus, nasal dorsum, sella turcica and clivus, neck, subglottis, and tonsil. Three cases presented with rapidly growing subcutaneous painless masses within 2 months, 1 case presented with hemoptysis, hoarseness and asthma for 28 days, 1 case presented with blood in the nasal discharge for 2 months, 1 case presented with headache for 1 month, and 1 case was found to have tonsillar neoplasms on physical examination for 3 days. CT was performed in 5 patients and the lesions showed soft tissue density shadows, and MRI was also performed in 2 of them, which showed T1 isointensity or T2 slightly long abnormal signal shadows. All patients underwent simple surgical resection of the mass. The patients were followed up for 13 months to 10 years, and none had recurrence. Conclusion: Primary NF in the head and neck is rare, with atypical clinical symptoms and imaging findings. Surgery is the mainstay of treatment for NF with good results.

[The efficacy and safety of Bruton tyrosine kinase inhibitors as monotherapy in the treatment of newly diagnosed patients with Waldenström macroglobulinemia].

Objective: To investigate the efficacy and safety of Bruton tyrosine kinase inhibitors (BTKi) ibrutinib or zanubrutinib monotherapy in newly diagnosed patients with Waldenström macroglobulinemia (WM) . Methods: The efficacy and adverse effects of 58 patients with newly diagnosed WM receiving BTKi monotherapy in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine were analyzed retrospectively from January 2018 to August 2022. Results: The response of 55 patients may be examined. Forty patients received ibrutinib monotherapy for a median of 15 months, with an overall response rate (ORR) of 85%, a main remission rate (MRR) of 70%, and a very good partial remission (VGPR) rate of 10%. Fifteen patients received zanubrutinib monotherapy for a median of 13 months, with an ORR of 93%, an MRR of 73%, and a VGPR rate of 0%. For various reasons, 10 patients were converted from ibrutinib to zanubrutinib. Ibrutinib treatment lasted an average of 7.5 months before conversion. The median duration of zanubrutinib therapy after conversion was 3.5 months. The ORRs before and after conversion were 90% and 100%, MRRs were 80% and 80%, and VGPR rates were 10% and 50%, respectively. After a median of 16 months, the 24-month progression-free survival (PFS) rate of patients who received both BTKi was 86%. PFS did not differ statistically across individuals with low, medium, and high-risk ISS scores (P=0.998). All of the patients survived. The most common side effects of BTKi were neutropenia and thrombocytopenia, which occurred in 12% and 10% of all patients, respectively. Ibrutinib accounts for 5% of atrial fibrillation, and zanubrutinib has a 7% risk of bleeding. Conclusions: In treating WM, ibrutinib or zanubrutinib provides good efficacy and tolerable adverse effects.

HMGB1 acts as an agent of host defense at the gut mucosal barrier.

Mucosal barriers provide the first line of defense between internal body surfaces and microbial threats from the outside world. 1 In the colon, the barrier consists of two layers of mucus and a single layer of tightly interconnected epithelial cells supported by connective tissue and immune cells. 2 Microbes colonize the loose, outer layer of colonic mucus, but are essentially excluded from the tight, epithelial-associated layer by host defenses. 3 The amount and composition of the mucus is calibrated based on microbial signals and loss of even a single component of this mixture can destabilize microbial biogeography and increase the risk of disease. 4-7 However, the specific components of mucus, their molecular microbial targets, and how they work to contain the gut microbiota are still largely unknown. Here we show that high mobility group box 1 (HMGB1), the prototypical damage-associated molecular pattern molecule (DAMP), acts as an agent of host mucosal defense in the colon. HMGB1 in colonic mucus targets an evolutionarily conserved amino acid sequence found in bacterial adhesins, including the well-characterized Enterobacteriaceae adhesin FimH. HMGB1 aggregates bacteria and blocks adhesin-carbohydrate interactions, inhibiting invasion through colonic mucus and adhesion to host cells. Exposure to HMGB1 also suppresses bacterial expression of FimH. In ulcerative colitis, HMGB1 mucosal defense is compromised, leading to tissue-adherent bacteria expressing FimH. Our results demonstrate a new, physiologic role for extracellular HMGB1 that refines its functions as a DAMP to include direct, virulence limiting effects on bacteria. The amino acid sequence targeted by HMGB1 appears to be broadly utilized by bacterial adhesins, critical for virulence, and differentially expressed by bacteria in commensal versus pathogenic states. These characteristics suggest that this amino acid sequence is a novel microbial virulence determinant and could be used to develop new approaches to diagnosis and treatment of bacterial disease that precisely identify and target virulent microbes.