OBJECTIVES: Metastasis is the primary cause of death in osteosarcoma, and current clinical treatments remain limited. BRD4, a key epigenetic regulator, has shown therapeutic promise in various cancers through its inhibition. However, the mechanistic role of BRD4 in osteosarcoma remains poorly understood. This study aims to elucidate the molecular mechanisms by which BRD4 regulate osteosarcoma progression and to explore novel therapeutic strategies. METHODS: Immunofluorescence was used to assess BRD4 expression levels in a tissue microarray containing 80 osteosarcoma samples from different patients. The Gene Expression Omnibus (GEO) dataset (GSE42352, containing survival data from 88 osteosarcoma patients) was downloaded to perform Kaplan-Meier survival analysis based on BRD4 gene expression levels. In vivo, an orthotopic intramedullary osteosarcoma model was established using HOS cells in C57 mice, followed by treatment with varying doses of the BRD4 inhibitor (+)-JQ1. Micro-CT, 3D reconstruction of bone tissue, and HE staining were employed to evaluate pathological changes in bone and intestinal lymph nodes. In vitro, cell viability was measured using the methyl thiazolyl tetrazolium (MTT) assay, while colony formation and Transwell assays assessed proliferative and invasive capacities. Chromatin-bound BRD4 was analyzed via co-immunoprecipitation combined with mass spectrometry (Co-IP/MS), and O-GlcNAc glycosylation sites and glycan chains of BRD4 were identified using Co-IP with Nano-LC MS/MS. Real-time PCR and Western blotting were used to analyze the relative mRNA and protein expression levels of target genes, respectively. RESULTS: BRD4 was positively expressed in 61.25% (49/80) of osteosarcoma tissues. Patients with high BRD4 expression exhibited significantly shorter survival times (P<0.05). In the orthotopic mouse model, intervention with (+)-JQ1, a potent and commonly used BETi, significantly inhibited tumor growth in vivo and reduced bone destruction (P<0.05). (+)-JQ1 treatment significantly suppressed the proliferation (P<0.001), invasion (P<0.001), and migration (P<0.05) of HOS cells. In osteosarcoma cells, BRD4 exhibited O-GlcNAc modifications at both N- and C- C-termini, particularly at Thr73, which is essential for protein stability. This modification also contributed to the activation of the EGFR tyrosine kinase inhibitor resistance pathway (KEGG Pathway: hsa01521). (+)-JQ1 treatment displaced BRD4 from enhancers and downregulated the transcription of pathway-related genes, such as EGFR and PDGFC, thereby suppressing the malignant behavior of osteosarcoma cells. CONCLUSIONS BRD4 promotes osteosarcoma progression via O-GlcNAc modification at Thr73 and plays a crucial role in tumor growth and metastasis.
Osteosarcoma/drug therapy* ; Humans ; Transcription Factors/metabolism* ; Animals ; Cell Cycle Proteins ; Mice ; Bone Neoplasms/drug therapy* ; Azepines/pharmacology* ; Cell Line, Tumor ; Cell Proliferation/drug effects* ; Triazoles/pharmacology* ; Mice, Inbred C57BL ; Nuclear Proteins/metabolism* ; Gene Expression Regulation, Neoplastic ; Male ; Bromodomain Containing Proteins

Objective To discuss the application of ultrasound multimodal diagnostic mode combined with ultrasonic precision interventional technology in differentiating the benign from malignant solid breast masses.Methods A total of 396 patients who underwent breast surgery at the Wuwei Cancer Hospital of China from May 2021 to July 2023 were enrolled in this study.Within 2 weeks before surgery,multimodal ultrasound examination(including two-dimensional ultrasound,color Doppler ultrasound,elastic ultrasound,breast three-dimensional ultrasound,and contrast-enhanced ultrasound)and ultrasound-guided needle biopsy were performed in all patients.The consistency between the results of multimodal ultrasound examination,ultrasound-guided needle biopsy,combination diagnosis and the postoperative pathological diagnoses was analyzed.Results Of the 396 patients with solid breast mass,malignant lesion was seen in 237(59.85％)and benign lesion was seen in 159(40.15％).The sensitivity of multimodal ultrasound in diagnosing benign and malignant solid breast masses was 77.64％(184/237),the specificity was 90.57％(144/159),the positive predictive value was 92.46％(184/199),the negative predictive value was 73.10％(144/197),the accuracy was 82.83％(328/396),and the consistency with the postoperative pathological diagnosis was 0.656.The sensitivity of ultrasound-guided needle puncture in diagnosing benign and malignant solid breast masses was 94.51％(224/237),the specificity was 100.00％(159/159),the positive predictive value was 100.00％(224/224),the negative predictive value was 92.44％(159/172),the accuracy was 96.72％(383/396),and the consistency with the postoperative pathological diagnosis was 0.933.The sensitivity of multimodal ultrasound combined with ultrasound-guided needle puncture in diagnosing benign and malignant solid breast masses was 100.00％(228/228),the specificity was 94.64％(159/168),the positive predictive value was 96.20％(228/237),the negative predictive value was 100.00％(159/159),the accuracy was 97.73％(387/396),and the consistency with the postoperative pathological diagnosis was 0.937.Conclusion The ultrasound multimodal diagnostic mode and ultrasonic precision interventional technology can be used in differentiating the benign from malignant solid breast masses with high diagnostic accuracy.

Objective: To investigate the characteristics of breakthrough cases of varicella in Haishu District, Ningbo City, Zhejiang Province from 2017 to 2022, so as to provide the evidence for varicella prevention and control. Methods: Information on reported cases of varicella and vaccination in Haishu District from 2017 to 2022 were collected through the China Infectious Disease Reporting Management Information System and the Immunization Program Information Management System of Zhejiang Province. The breakthrough cases who had received 1 or 2 doses of varicella vaccine 42 days before the onset of disease were selected. The distribution of sex, age, region and the interval between onset and last immunization were descriptively analyzed. Results: A total of 1 563 varicella cases were reported from 2017 to 2022. There were 928 breakthrough cases (59.37%), of which 660 cases with 1-dose immunization history (42.23%) and 268 cases with 2-dose immunization history (17.15%). The proportion of 1-dose breakthrough cases in total annual cases showed a decreasing trend (P<0.05) from 2017 to 2022, while there was no significant tendency on the proportion of cases with 2-dose immunization history (P>0.05). There were 392 males (59.39%) and 268 females (40.61%) with 1-dose immunization history. There were 150 males (55.97%) and 118 females (44.03%) with 2-dose immunization history. Breakthrough cases with 1- and 2- dose immunization history had an median age of 13.00 (interquartile range, 4.00) and 9.00 (4.00) years, respectively, and the difference was statistically significant (P<0.05). The regional distribution of the breakthrough cases with 1- and 2- dose immunization history was mainly in the rural-urban fringe, with 319 cases (48.33%) and 137 cases (51.12%), respectively, and the difference was statistically significant (P<0.05). The median interval between onset and last immunization was 12.00 (interquartile range, 3.00) and 4.00 (3.00) years, respectively, and the difference was statistically significant (P<0.05). Conclusions The breakthrough cases of varicella in Haishu District from 2017 to 2022 were mainly cases with 1-dose immunization history, males, and residents in rural-urban fringe. The age was older and the interval between onset and the last immunization was longer in cases with 1-dose immunization history than in cases with 2-dose immunization history.

Objective:To evaluate clinical prognosis and prognostic factors of patients with early stage (Ⅰ stage) and locally advanced (Ⅱ/Ⅲ stage) lung cancer treated with carbon ion radiotherapy (CIRT).Methods:Clinical data, treatment, adverse reactions, survival and so on of 54 lung cancer patients who received CIRT and follow-up in the Heavy Ion Center of Wuwei Cancer Hospital of Gansu Province from March 2020 to September 2022 were retrospectively analyzed. The survival curve was plotted using Kaplan-Meier method. Difference tests were performed using log-rank test. Logistic regression analysis was used to identify prognostic factors.Results:According to inclusion and exclusion criteria, 54 patients were enrolled in the study, including 10 patients with early stage lung cancer and 44 patients with locally advanced lung cancer. The median follow-up time for 10 patients with early stage lung cancer was 11.0 (6.75, 17.25) months, and the median dose of irradiation was 60 Gy [relative biological effect (RBE)]. Upon the last follow-up, 3 patients had complete response (CR) and 3 patients had partial response (PR). Four patients had stable disease (SD) and no progressive disease (PD). The 1-year and 2-year local control rates (LCR), progression-free survival (PFS) rates and overall survival (OS) rates were 100%. During treatment and follow-up, 2 patients developed grade 1 radiation pneumonia, 1 case of grade 2 radiation pneumonia, 1 case of chest wall injury (chest wall pain), and there were no adverse reactions greater than grade 2. The median follow-up time of 44 patients with locally advanced stage was 12.5 (4.25, 21.75) months, and the median irradiation dose was 72 Gy (RBE). Thirty-two (73%) patients received concurrent chemotherapy during treatment, 20 (45%) patients received sequential chemotherapy after treatment, 14 (32%) patients received immune maintenance therapy and 3 (7%) patients obtained PD and received targeted drugs. Upon the last follow-up, 3 (7%) patients had CR, 17 (39%) patients had PR, 19 (43%) patients obtained SD, and 5 (11%) patients had PD. The 1-year and 2-year LCR were 96.0% and 87.3%, 90.9% and 84.1% for the 1-year and 2-year PFS rates, and 93.2% and 86.4% for the 1-year and 2-year OS rates, respectively. The median OS and PFS of patients were not reached. Multivariate logistic regression analysis showed that maintenance therapy after radiotherapy ( P=0.027) and clinical target volume (CTV) irradiation volume ( P=0.028) were the factors affecting PFS. Simultaneous chemoradiotherapy ( P=0.042) and maintenance therapy after radiotherapy ( P=0.020) were the factors affecting OS. And gross tumor volume (GTV) ≥215 ml ( P=0.068) might be an independent risk factor for grade 2 and above radiation pneumonia. Conclusions:The domestic carbon ion system has definite clinical effect and controllable toxic and side effects in the treatment of early stage and locally advanced lung cancer. The combination of synchronous chemotherapy and further maintenance treatment can significantly improve clinical prognosis of patients without significantly increasing the risk of toxic and side effects.

ObjectiveTo evaluate the reliability and validity of the Dampness Syndrome Scale of Chinese Medicine （DSSCM） among patients with persistent asthma， and to explore the correlation between dampness syndrome and clinical characteristics of persistent asthma. MethodsA cross-sectional survey was conducted. Basic information， examination results， DSSCM， Asthma Control Test （ACT）， Generalized Anxiety Disorder-7 （GAD-7）， and Patient Health Questionnaire-9 （PHQ-9） scores were collected from 206 patients with persistent asthma to evaluate the reliability and validity of DSSCM and to explore the correlation between dampness syndrome and clinical characteristics. ResultsThe mean score of DSSCM among 206 patients was 14.59 ± 10.53. The overall Cronbach α coefficient and Spearman-Brown split-half reliability coefficient of the scale were both greater than 0.8， and the success rate of scale convergent and discriminant validity calibration were greater than 80%. The confirmatory factor analysis showed that the χ²/df was 2.309， and the root mean square error of approximation （RMSEA） was 0.08； the root mean square residual （RMR） was 0.049， whereas the comparative fit index （CFI）， the goodness of fit index （GFI）， the adjusted goodness of fit index （AGFI）， the normed fit index （NFI） and the incremental fit index （IFI） were less than 0.9. Correlation analysis showed that DSSCM scores were positively correlated with disease duration， GAD-7 scores， and PHQ-9 scores （P＜0.05）， and negatively correlated with ACT scores （P＜0.01）. The DSSCM scores were significantly different between patients with different disease severity （H = 10.92， P = 0.01）， and the DSSCM scores of allergic patients were higher than those of non-allergic patients （Z = -4.19， P＜0.001）. ConclusionDSSCM has acceptable reliability and validity for patients with persistent asthma. The scores of DSSCM correlated with the disease duration， ACT score， GAD-7 score， PHQ-9 score， disease severity and allergic status of persistent asthmatics.

Objective:To investigate the clinical characteristics of psoriasis and status quo of medical care for patients in county areas of China.Methods:This study was a cross-sectional investigation. Based on the “Qianxian Wuyin” Project (a national project for upgrating ability for psoriasis care at county level), an online questionnaire survey was conducted in the dermatology departments of 459 county hospitals in 404 pilot administrative counties across China from February to June 2023. The questionnaire included demographic information of patients (gender, ethnicity, age, place of residence, education, marital status), and clinical characteristics of psoriasis (disease course, type, comorbidities, body surface area (BSA) and previous treatment. The Dermatology Life Quality Index (DLQI) and Psoriasis Area and Severity Index (PASI) were applied for assessing the quality of life and disease severity, and completed by patients or guardian and doctors, respectively.Results:A total of 16 935 patients completed the questionnaire. The age of patients was 1-102(44.17±11.58)years, and 71.0% (12 036/16 935) were 30-59 years old. The ratio of male to female was 2.21∶1; 24.3%(4 117/16 935) of patients had high school education; there were 9 940 patients(58.7%) with previous or current smoking and/or alcohol use; 42.8%(7 218/16 855) of patients had a disease course of 1-5 years. There were 15 630 patients(92.3%) with DLQI≥10, 8 346 patients(49.7%) with PASI≥10, 15 017 patients(89.2%) with BSA≥10%. The plaque type was the most common disease type ( n=14 965, 88.7%), and spotting type ranked the second ( n=1 141, 6.8%). The most common initial site was the trunk ( n=12 309, 72.9%). Among the comorbidities, hypertension was the most common one ( n=1 681, 10.0%). There were 7 650 reports of treatment response to conventional topical drug therapy and 3 112 reports of treatment response to systemic drug therapy, with 6 269 (81.9%) and 2 493 (80.1%) reporting poor or no response, respectively. Conclusions:The survey shows that in the county areas of China, the majority of psoriasis patients are severe patients with short course of disease, plaque type is the most common type, and hypertension is the most common comorbidity; and the conventional treatment is less effective for most patients.

Overweight/obesity is an important risk factor for gestational diabetes mellitus (GDM). Overweight/obesity and GDM can affect maternal and infants to varying degrees. Reducing the incidence of GDM in overweight/obese people by early intervention is imminent. Current researches on interventions mainly focus on exercise and diet, remote interventions with electronic devices, nutritional supplements, medications, and weight loss surgery. Exercise and diet interventions are the most significant means to reduce the incidence of GDM, and their effectiveness can be affected by the timing, duration, and intervention methods. Maternal compliance is also an essential influencing factor. The efficacy of remote interventions by electronic devices, various nutritional supplements, and drug interventions remains controversial. Weight loss surgery may offer potential benefits but may have some implications on maternal nutritional status and fetal growth and development. Pre-pregnancy weight loss and prevention of excessive weight gain during pregnancy are key initiatives to reduce the incidence of GDM.

Objective To explore the surgical techniques and experiences in the treatment of Peyronie's disease using electric rotary grinding of tunica albuginea plaques followed by graft repair with tunica vaginalis or bovine pericardium under a microscope.Methods A retrospective cohort study was performed on 21 patients with severe penile curvature due to Peyronie's disease admitted in our department from January 2021 and May 2023.All of them underwent microscopic electric rotary grinding of tunica albuginea plaques followed by graft repair surgery.Short-term and mid-term outcomes were observed.Results For the 21 enrolled patients,12(57.1％)underwent repair using unilateral tunica vaginalis,3(14.3％)underwent repair using bilateral tunica vaginalis,and 6(28.6％)underwent repair using bovine pericardium.The operation time was 170～385(average 272±69)min,and the average postoperative follow-up duration was3～24(average 12.1±7.2)months.Seventeen patients(81.0％)had their penis fully extended,while 4 patients(19.0％)had slight dorsal curvature,which was less than 10°.All patients were satisfied with the shape of their penis during erection.The change in penile length was+(1.1±0.5)cm,and 9 patients(42.9％)reported early postoperative penile head numbness,which spontaneously resolved after an average duration of 3.2±1.9 months.All patients were able to successfully engage in sexual activity,with an average penile erection hardness score of 3.8.The overall satisfaction rate was 90.5％post operatively.Conclusion Meticulous dissection of Buck's fascia under a microscope is advantageous in reliably preserving dorsal neurovascular structures.Application of an electric grinding drill for plaque ablation allows for more precise release of the tunica.Graft repair of tunica albuginea defects with testicular tunica vaginalis or bovine pericardium for Peyronie's disease yields overall favorable clinical outcomes and obtains high satisfaction.It is worthy of promotion in clinical practice.

Objective To compare the preparation efficiency of mouse pox and mouse hepatitis antibodies between two substrains of BALB/c and Big-BALB/c (B-BALB/c) mice, and to provide a theoretical basis and reference for the selection of laboratory animals in the preparation of monoclonal antibodies inducedin vivo through hybridoma.Methods Individuals weighing more than 5% of the weight of normal animals at 4 weeks of age (the criterion for late selection is more than 10%) were selected from a population of conventionally bred BALB/c mice and bred individually, and a subline of B-BALB/c mice was prepared after 10 generations of selection. A total of 40 BALB/c mice and 40 B-BALB/c mice aged 10 to 11 weeks, half male and half female, were selected and inoculated with the mousepox monoclonal antibody hybridoma cell line G23 or the murine hepatitis monoclonal antibody hybridoma cell line Y15 pre-treated with liquid paraffin, respectively. Mice ascites containing monoclonal antibodies were obtained by in vivo induction. The antibody titer was tested by indirect ELISA. The mice were grouped based on the sub-strains, gender and inoculation type of hybridoma to analyze the ascites production, antibody titer and antibody production, and to evaluate the antibody preparation efficiency of the two BALB/c mouse sub-strains.ResultsAfter 10 generations of breeding, the body weight of 10-week-old male and female B-BALB/c mice increased by 22.3% and 12.8%, respectively, compared with BALB/c mice of the same age. Compared with BALB/c mice, B-BALB/c mice had better tolerance and adaptation to secondary ascites collection. Compared with BALB/c mice, the ascites production and antibody titer during the preparation of antibodies in B-BALB/c mice were significantly increased, especially in the hybridoma cell line G23 vaccination group (both P＜0.000 1) . After inoculation with the hybridoma cell lines G23 or Y15, the average antibody production of B-BALB/c mice (14.99×10⁴ U and 33.22×10⁴ U) was higher than that of BALB/c mice (5.33×10⁴ U and 19.31×10⁴ U) (both P＜0.01). After inoculation with hybridoma cell line G23, the average antibody production per unit body weight of B-BALB/c mice (0.55×10⁴ U/g) was higher than that of BALB/c mice (0.23×10⁴ U/g) (P＜0.000 1). And the antibody production per unit body weight of female B-BALB/c or BALB/c mice was higher than that of male B-BALB/c or BALB/c mice (bothP＜0.01).Conclusion B-BALB/c mice can be used as an alternative to BALB/c mice in the in vivo induction of monoclonal antibody preparation, which can achieve the purpose of reducing the number of experimental animals used, lowering the labor cost, and improving the efficiency of antibody preparation.

OBJECTIVE: To investigate the accuracy, safety, and short-term effectiveness of a domestic robot-assisted system in total knee arthroplasty (TKA) by a multicenter randomized controlled trial. METHODS: Between December 2021 and February 2023, 138 patients with knee osteoarthritis who received TKA in 5 clinical centers were prospectively collected, and 134 patients met the inclusion criteria were randomly assigned to either a trial group ( n=68) or a control group ( n=66). Seven patients had lost follow-up and missing data, so they were excluded and the remaining 127 patients were included for analysis, including 66 patients in the trial group and 61 patients in the control group. There was no significant difference ( P>0.05) in gender, age, body mass index, side, duration of osteoarthritis, Kellgren-Lawrence grading, preoperative Knee Society Score (KSS) and Western Ontario and McMaster University Osteoarthritis Index (WOMAC) score between the two groups. The trial group completed the TKA by domestic robot-assisted osteotomy according to the preoperative CT-based surgical planning. The control group was performed by traditional osteotomy plate combined with soft tissue release. Total operation time, osteotomy time of femoral/tibial side, intraoperative blood loss, and postoperative complications were recorded and compared between the two groups. The radiographs were taken at 5 and 90 days after operation, and hip-knee-ankle angle (HKA), lateral distal angle of femur (LDFA), and posterior tibial slope (PTS) were measured. The difference between the measured values of the above indexes at two time points after operation and the preoperative planning target values was calculated, and the absolute value (absolute error) was taken for comparison between the two groups. The postoperative recovery of lower limb alignment was judged and the accuracy was calculated. KSS score and WOMAC score were used to evaluate the knee joint function of patients before operation and at 90 days after operation. The improvement rates of KSS score and WOMAC score were calculated. The function, stability, and convenience of the robot-assisted system were evaluated by the surgeons. RESULTS: The total operation time and femoral osteotomy time of the trial group were significantly longer than those of the control group ( P<0.05). There was no significant difference in the tibial osteotomy time and the amount of intraoperative blood loss between the two groups ( P>0.05). The incisions of both groups healed by first intention after operation, and there was no infection around the prosthesis. Nine patients in the trial group and 8 in the control group developed lower extremity vascular thrombosis, all of which were calf intermuscular venous thrombosis, and there was no significant difference in the incidence of complications ( P>0.05). All patients were followed up 90 days. There was no significant difference in KSS score and WOMAC score between the two groups at 90 days after operation ( P>0.05). There was significant difference in the improvement rate of KSS score between the two groups ( P<0.05), while there was no significant difference in the improvement rate of WOMAC score between the two groups ( P>0.05). Radiological results showed that the absolute errors of HKA and LDFA in the trial group were significantly smaller than those in the control group at 5 and 90 days after operation ( P<0.05), and the recovery accuracy of lower limb alignment was significantly higher than that in control group ( P<0.05). The absolute error of PTS in the trial group was significantly smaller than that in the control group at 5 days after operation ( P<0.05), but there was no significant difference at 90 days between the two groups ( P>0.05). The functional satisfaction rate of the robot-assisted system was 98.5% (65/66), and the satisfaction rates of stability and convenience were 100% (66/66). CONCLUSION Domestic robot-assisted TKA is a safe and effective surgical treatment for knee osteoarthritis, which can achieve favorable lower limb alignment reconstruction, precise implant of prosthesis, and satisfactory functional recovery.
Humans ; Arthroplasty, Replacement, Knee/methods* ; Osteoarthritis, Knee/surgery* ; Blood Loss, Surgical ; Robotics ; Knee Joint/surgery* ; Knee Prosthesis ; Retrospective Studies