OBJECTIVE: Circadian rhythm disruption (CRD) is a risk factor that correlates with poor prognosis across multiple tumor types, including hepatocellular carcinoma (HCC). However, its mechanism remains unclear. This study aimed to define HCC subtypes based on CRD and explore their individual heterogeneity. METHODS: To quantify CRD, the HCC CRD score (HCCcrds) was developed. Using machine learning algorithms, we identified CRD module genes and defined CRD-related HCC subtypes in The Cancer Genome Atlas liver HCC cohort (n = 369), and the robustness of this method was validated. Furthermore, we used bioinformatics tools to investigate the cellular heterogeneity across these CRD subtypes. RESULTS: We defined three distinct HCC subtypes that exhibit significant heterogeneity in prognosis. The CRD-related subtype with high HCCcrds was significantly correlated with worse prognosis, higher pathological grade, and advanced clinical stages, while the CRD-related subtype with low HCCcrds had better clinical outcomes. We also identified novel biomarkers for each subtype, such as nicotinamide n-methyltransferase and myristoylated alanine-rich protein kinase C substrate-like 1. CONCLUSION We classify the HCC patients into three distinct groups based on circadian rhythm and identify their specific biomarkers. Within these groups greater HCCcrds was associated with worse prognosis. This approach has the potential to improve prediction of an individual's prognosis, guide precision treatments, and assist clinical decision making for HCC patients. Please cite this article as: Li XJ, Chang L, Mi Y, Zhang G, Zhu SS, Zhang YX, et al. Integrated-omics analysis defines subtypes of hepatocellular carcinoma based on circadian rhythm. J Integr Med. 2025; 23(4): 445-456.
Humans ; Carcinoma, Hepatocellular/pathology* ; Liver Neoplasms/pathology* ; Circadian Rhythm/genetics* ; Prognosis ; Male ; Female ; Biomarkers, Tumor/genetics* ; Middle Aged ; Machine Learning ; Computational Biology

Endotoxins are common exogenous pyrogens. Excessive endotoxins in medical devices and injections can lead to serious consequences such as sepsis, septic shock, and even death. Therefore, endotoxin detection plays a crucial role in medical, pharmaceutical, and food sectors. The wide application of Limulus amebocyte lysate (LAL) has led to a sharp decline in the number of horseshoe crabs. Moreover, the LAL assay has limitations such as interbatch variations and difficulty in quantification. The recombinant factor C (rFC) assay is stable between batches, highly sensitive, and capable of quantitation, and thus it can be used as an alternative for the LAL assay. However, the high cost and complex procedures involved in producing recombinant factor C have limited the widespread application of this method. In order to simplify the preparation and reduce the production cost of recombinant factor C, this study focuses on the production of recombinant factor C based on the baculovirus expression system. Multiple measures such as a high-yield and anti-apoptotic vector qBac-IIIG, the optimal signal peptide, and the optimized codon were used to reach the goal of endotoxin detection with cell supernatant. This method simplifies the steps of protein purification. The sensitivity of the supernatant reached 0.05 EU/mL in a 1-L fermentation system, and 500 000 detecting reactions can be supported per liter of fermentation broth. This study increases the yield and activity of recombinant factor C, simplifies the procedures of protein purification, and reduces the cost, laying a foundation for the promotion and application of recombinant factor C in endotoxin detection.
Animals ; Recombinant Proteins/genetics* ; Horseshoe Crabs/chemistry* ; Baculoviridae/metabolism* ; Endotoxins/analysis* ; Protein C/biosynthesis* ; Genetic Vectors/genetics* ; Arthropod Proteins/genetics* ; Enzyme Precursors ; Serine Endopeptidases

Objective To investigate the role of miR-125b on hepatic angiogenesis,with the hope of providing new targets for the prevention and treatment of liver fibrosis.Methods The human hepatic sinusoidal endothelial cells were transfected with miR-125b mimics and inhibitors,and the mRNA and protein expression of vascular endotheli-al growth factor(VEGF),cluster of differentiation antigens 31(CD31),von Willebrand factor(vWF),collagenⅣ,and laminin(LN)were detected by qRT-PCR and ELISA,and the expression of nitric oxide(NO)was detec-ted by fluorescent probe,scanning electron microscopy detected the alteration of the window holes on the surface of human hepatic sinusoidal endothelial cells,angiogenesis assay was performed to observe the neovascularization of each group,and dual luciferase reporter gene assay was performed to validate the targeting relationship between miR-125b and VEGF.Results qRT-PCR and ELISA showed that compared with the negative control group,the mRNA and protein expressions of VEGF,CD31,vWF,Collagen Ⅳ,and LN significantly decreased after miR-125b mimic transfection(P＜0.05),while the mRNA and protein expressions of VEGF,CD31,vWF,CollagenⅣ,and LN were significantly increased after transfection with miR-125 b mimics(P＜0.05);fluorescent probe detection showed that compared with the negative control group,the average fluorescence of intensity expression NO decreased significantly(P＜0.05),while the average fluorescence intensity expression of NO increased significant-ly after miR-125b inhibitor transfection(P＜0.05);the number of fenestrations on the surface of human liver sinu-soidal endothelial cells significantly increased after miR-125b mimic transfection(P＜0.05),while the number of fenestrations on the surface of human liver sinusoidal endothelial cells decreased significantly after miR-125 b inhibi-tor transfection(P＜0.05);angiogenesis assay showed that compared with the negative control group,the number of angiogenesis significantly decreased after miR-125b mimic transfection(P＜0.05),while the number of angio-genesis significantly increased after miR-125b inhibitor transfection(P＜0.05);dual luciferase reporter gene assay showed that compared with negative control group,the expression of relative fluorescence intensity after transfection of miR-125b mimics in VEGF wild-typ significantly decreased(P＜0.05),while the expression of relative fluores-cence intensity after transfection of miR-125b mimics in VEGF mutant significantly decreased(P＞0.05).Con-clusion miR-125b can inhibit liver angiogenesis and thus play an anti-fibrosis role,which can provide a new ref-erence for the prevention and treatment of chronic liver disease and the development of new drugs.

Objective:To assess the risk of noise-induced hearing loss in workers from a petrochemical plant.Methods:In October 2020, 488 male workers exposed to noise in a petrochemical plant in Guangdong Province were selected by cluster sampling. Acoustics-Estimation of Noise-Induced Hearing Loss (ISO 1999: 2013) was used to assess the risk of noise-induced hearing loss of workers, and individual fit testing was used to evaluate the sound attenuation obtained by the workers. The risk assessment results and fitness test results of workers with different hearing levels were compared.Results:The average noise exposure equivalent sound level of the workers in the petrochemical plant was 86.7 dB (A) . The median of PARs (personal attenuation ratings) was 16 (4, 23) dB. There were statistically significant differences in age and service years among workers with different hearing results ( P<0.05) , but no statistically significant differences in noise intensity and PARs ( P>0.05) . According to risk assessment results of ISO 1999: 2013, the current risk of high-frequency hearing loss in 488 workers were negligible risk and acceptable risk. The risk of noise-induced deafness weredivided into three levels: negligible risk in 452 workers (92.7%) , medium risk in 27 workers (5.5%) and high risk in 9 workers (1.8%) . The risk of high-frequency hearing loss in next 5 to 15 years for workers with noise exposure level of >94 to 97 dB and >97 dB or above would be medium risk or above. The risk of noise-induced deafness in next 5 to 15 years for workers exposed to noise withlevel of 91 to 94 dB would be medium risk or above. Conclusion:The risk of noise-induced hearing loss in workers from the petrochemical plant is high in next 5 to 15 years, and noise prevention and control measures need to be strengthened. ISO1999: 2013 assessment method may underestimate the risk of hearing loss among workers.

Objective:To assess the risk of noise-induced hearing loss in workers from a petrochemical plant.Methods:In October 2020, 488 male workers exposed to noise in a petrochemical plant in Guangdong Province were selected by cluster sampling. Acoustics-Estimation of Noise-Induced Hearing Loss (ISO 1999: 2013) was used to assess the risk of noise-induced hearing loss of workers, and individual fit testing was used to evaluate the sound attenuation obtained by the workers. The risk assessment results and fitness test results of workers with different hearing levels were compared.Results:The average noise exposure equivalent sound level of the workers in the petrochemical plant was 86.7 dB (A) . The median of PARs (personal attenuation ratings) was 16 (4, 23) dB. There were statistically significant differences in age and service years among workers with different hearing results ( P<0.05) , but no statistically significant differences in noise intensity and PARs ( P>0.05) . According to risk assessment results of ISO 1999: 2013, the current risk of high-frequency hearing loss in 488 workers were negligible risk and acceptable risk. The risk of noise-induced deafness weredivided into three levels: negligible risk in 452 workers (92.7%) , medium risk in 27 workers (5.5%) and high risk in 9 workers (1.8%) . The risk of high-frequency hearing loss in next 5 to 15 years for workers with noise exposure level of >94 to 97 dB and >97 dB or above would be medium risk or above. The risk of noise-induced deafness in next 5 to 15 years for workers exposed to noise withlevel of 91 to 94 dB would be medium risk or above. Conclusion:The risk of noise-induced hearing loss in workers from the petrochemical plant is high in next 5 to 15 years, and noise prevention and control measures need to be strengthened. ISO1999: 2013 assessment method may underestimate the risk of hearing loss among workers.

Objective:To investigate the application effects of self-made simple vacuum sealing drainage (VSD) device in the postoperative treatment of sural neurocutaneous flap transplantation in the foot and ankle.Methods:From January 2017 to January 2019, 36 patients with foot and ankle skin defects and bone exposure admitted to People′s Hospital of Xinjiang Uygur Autonomous Region met the inclusion criteria, and a retrospective cohort study was conducted. According to the bandaging method of the operative area, simple negative pressure group and antibacterial dressing group were both allocated with 18 patients, with 12 males and 6 females in the former group, 14 males and 4 females in the latter group, aged (41.6±2.8) and (42.3±2.6) years, respectively. Patients in the two groups all received sural neurocutaneous flap transplantation. Patients in antibacterial dressing group received nano silver antibacterial dressing change in the operative area, and the dressing was changed once every 3 days. In simple negative pressure group, the operative area was sealed with a simple VSD device made of gauze, silicone sputum suction tube with holes cut out, and biological permeable membrane, etc., which was connected with the wall central negative pressure suction system for continuous VSD treatment of -40.0 to -16.6 kPa. The negative pressure material was changed once every 5 days. The number of dressing change, the pain score evaluated by Numeric Rating Scale during each dressing change, the cost of dressing change, and the degree of flap swelling evaluated on the 3rd and 5th day after surgery were recorded, and the flap survival was observed. Data were statistically analyzed with independent sample t test, Wilcoxon rank sum test, and chi-square test. Results:The number of dressing change of patients in simple negative pressure group was (3.4±0.5) times, which was significantly less than (7.0±0.8) times in antibacterial dressing group ( t=15.338, P<0.01). The pain score during dressing change of patients in simple negative pressure group was (4.3±0.8) points, which was significantly lower than (6.8±0.7) points in antibacterial dressing group ( t=10.168, P<0.01). The cost of dressing change of patients was similar between the two groups. On the 3rd and 5th day after surgery, the degrees of flap swelling of patients in simple negative pressure group were significantly superior to those in antibacterial dressing group ( Z=4.448, 2.395, P<0.05 or P<0.01). The flap survival of patients in simple negative pressure group was significantly superior to that in antibacterial dressing group ( χ2=4.500, P<0.05). Conclusions:Compared with the traditional dressing bandage, the self-made simple VSD device used after sural neurocutaneous flap transplantation can reduce the frequency of dressing change, relieve the pain of dressing change and the swelling of flap, and promote the flap survival, which is worth popularizing and applying in clinic.

Objective To understand the current status of synthetic drug use and associated factors among men who have sex with men (MSM) at high risk of HIV infection.Methods A crosssectional study was conducted in Guangzhou,Wuxi and Shenzhen through January to August 2017.MSM at high risk of HIV infection were recruited from attendants at the health clinics,through internet advertisement and snowball sampling method.Eligible MSM were the ones who had met the following criteria:≥ 18 years old,having either 2 male sex partners,or condomless anal sex with a casual male sex partner,or a STD historg during the past 6 months.Data regarding demographic,sexual behavior,HIV/STD testing history and previous drug use were collected by sely-administered questionnaires.Blood was also drawn for HIV testing.Results Of the 603 MSM at high risk of HIV,25.5％ (154/603,95％CI:22.0％-29.0％) of them had used drugs in the past 6 months,including 29.1％ (88/302,95％CI:24.0％-34.3％) in Guangzhou,26.3％ (40/152,95％CI:19.2％-33.4％)in Shenzhen,and 17.4％ (26/149,95％CI:11.3％-23.6％) in Wuxi.'Rush'(85.1％,131/154) was the most commonly used drug.Results from the multivariable logistic regression revealed that drug use was associated with the following factors:having multiple casual sexual partners (aOR=3.24,95％CI:1.29-7.43) and alcohol use (aOR=1.78,95％CI:1.12-2.80) in the past 6 months after adjusting for age,education and monthly income.Conclusions Drug use seemed common among MSM population at high risk of HIV infection and was associated with higher risk behavior that leading to HIV infection.Synthetic drug use among MSM deserved more attention.

Objective To discuss the treatment program and experience of " 4P" theory in the treatment of severe infection wounds of perineum.Methods Retrospective analysis of 123 cases of severe infection wounds of perineum in our hospital from March 2016 to February 2018.Following the "4P" theory (T wound debridement-preparation,I prevention and treatment of infection-protection,M dry-wet balanceprogress,E wound edge management-promotion),personalized treatment program was implemented to ensure the normal physiological activities of patients while closing the wound and promoting wound healing.Results There was no death and no serious complications.All the patients were discharged from the hospital after the wound healed well.Wound healing time was (10.37 ±3.25)days.The follow-up time was 2 to 10 months,the median follow-up was 6 months.The recurrence occurred in 3 case and chronic pain in 5 cases during the follow-up.Conclusions According to the main factors affecting wound healing,"4P Theory" can protect the wound to the maximum extent and is safe and effective in the treatment of severe infection of perineal wound.

Objective: To explore the risk factors of diabetic foot ulcer (DFU) in diabetic patients of Uyghur nationality and Han nationality in the Xinjiang Uygur Autonomous Region. Methods: Clinical data of 640 diabetic patients admitted to our ward from January 2015 to November 2016, conforming to the study criteria, were retrospectively analyzed. Patients were divided into DFU group (n=403) and non-DFU group (n=237) according to whether DFU occurred or not. The data of gender, age, nationality, body mass index (BMI), smoking, drinking, binge eating, triglyceride (TG), total cholesterol (TC), and high-density lipoprotein (HDL) of patients between two groups were compared with chi-square test and t test. Indexes with statistically significant differences between two groups were selected, and they were processed with non-conditional multivariate logistic regression analysis to screen the independent risk factors of DFU. The possible risk factors of DFU of patients of Uyghur nationality and Han nationality were further processed with non-conditional multivariate logistic regression analysis respectively to screen the independent risk factors of DFU of patients of Uyghur nationality and Han nationality. Results: (1) There were no statistically significant differences in gender, age, TC, and HDL of patients between two groups (with χ²=0.149, t values respectively 1.163, 1.033, and 1.026, P values above 0.05). There were statistically significant differences in nationality, BMI, smoking, drinking, binge eating, and TG of patients between two groups (with χ² values from 4.778 to 13.694, t values respectively 4.703 and 4.237, P<0.05 or P<0.01). (2) Nationality, BMI, smoking, drinking, binge eating, and TG were the independent risk factors of DFU(with odds ratios respectively 1.488, 1.527, 1.736, 1.738, 1.382, and 1.648, 95% confidence intervals respectively 1.315-3.175, 1.488-4.393, 1.834-4.675, 1.474-2.695, 1.342-4.678, and 1.105-6.747, P values below 0.05). (3) Smoking, drinking, binge eating, and TG were the independent risk factors of DFU in diabetic patients of Uyghur nationality (with odds ratios respectively 1.673, 1.387, 1.328, and 1.486, 95% confidence intervals respectively 1.384-1.765, 1.414-1.659, 1.423-1.687, and 1.150-1.670, P values below 0.05). BMI, smoking, and drinking were the independent risk factors of DFU in diabetic patients of Han nationality (with odds ratios respectively 2.442, 1.604, and 1.251, 95% confidence intervals respectively 2.223-2.699, 1.268-2.028, and 1.164-1.344, P<0.05 or P<0.01). Conclusions Smoking, drinking, binge eating, and TG were the independent risk factors of DFU in diabetic patients of Uyghur nationality. BMI, smoking, and drinking were the independent risk factors of DFU in diabetic patients of Han nationality.

Objective To explore the effect of salmon calcitonin on osteoporosis induced by spinal cord injury. Methods 100 patients with osteoporosis induced by spinal cord injury from September 2011 to September 2014 in our department were included. They were randomly divided into control group (n=50) and observation group (n=50). The control group received vitamin D3 only, while the observation group received vitamin D3 combined with salmon calcitonin on the basis of rehabilitation physiotherapy, for 6 months. Visual Analogue Scale (VAS) of pain was evaluated in different periods. The bone mineral density (BMD) of lumbar spine and femoral neck, the parathyroid hormone (PTH), bone gla protein (BGP) and 1,25- dihydroxy vitamin D3 (1,25-(OH)2D3) were tested and recorded. Results The VAS score was lower in the observation group than in the control group 1, 2, 3 and 6 months after treatment (P<0.001). The BMD of lumbar spine and femoral neck was significantly higher, the PTH and BGP were significantly lower and the 1,25-(OH)2D3 was significantly higher in the observation group than in the control group after treatment (P<0.001). Conclusion Combination of salmon calcitonin can effectively reduce the bone pain and improve the BMD in patients with osteoporosis induced by spinal cord injury.