Over the past two decades, genome-wide association study(GWAS) has identified numerous genetic variants and loci associated with heritable diseases. With the gradual maturation and saturation of GWAS methodologies, transcriptome-wide association study(TWAS) offers a novel perspective by linkinggenetic phenotypes to gene expression levels. By integrating TWAS with other multi-omics analyses, researchers can gain a deeper understanding of heritable diseases. This article provides an overview of recent groundbreaking and representative TWAS methods and tools, analyzes their strengths and limitations, and discusses future trends in TWAS development.

Objective: To investigate the efficacy of magnesium-strontium co-doped hydroxyapatite mineralized collagen (MSHA/Col) in improving the bone repair microenvironment and enhancing bone regeneration capacity, providing a strategy to address the insufficient biomimetic composition and limited bioactivity of traditional hydroxyapatite mineralized collagen (HA/Col) scaffolds. Methods: A high-molecular-weight polyacrylic acid-stabilized amorphous calcium magnesium strontium phosphate precursor (HPAA/ACMSP) was prepared. Its morphology and elemental distribution were characterized by high-resolution transmission electron microscopy (TEM) and energy-dispersive spectroscopy. Recombinant collagen sponge blocks were immersed in the HPAA/ACMSP mineralization solution. Magnesium-strontium co-doped hydroxyapatite was induced to deposit within collagen fibers (experimental group: MSHA/Col; control group: HA/Col). The morphological characteristics of MSHA/Col were observed using scanning electron microscopy (SEM). Its crystal structure and chemical composition were analyzed by X-ray diffraction and Fourier transform infrared spectroscopy, respectively. The mineral phase content was evaluated by thermogravimetric analysis. The scaffold's porosity, ion release, and in vitro degradation performance were also determined. For cytological experiments, CCK-8 assay, live/dead cell staining, alkaline phosphatase staining, alizarin red S staining, RT-qPCR, and western blotting were used to evaluate the effects of the MSHA/Col scaffold on the proliferation, viability, early osteogenic differentiation activity, late mineralization capacity, and gene and protein expression levels of key osteogenic markers [runt-related transcription factor 2 (Runx2), collagen type Ⅰ (Col-Ⅰ), osteopontin (Opn), and osteocalcin (Ocn)] in mouse embryonic osteoblast precursor cells (MC3T3-E1). Results: HPAA/ACMSP appeared as amorphous spherical nanoparticles under TEM, with energy spectrum analysis showing uniform distribution of carbon, oxygen, calcium, phosphorus, magnesium, and strontium elements. SEM results of MSHA/Col indicated successful complete intrafibrillar mineralization. Elemental analysis showed the mass fractions of magnesium and strontium were 0.72% (matching the magnesium content in natural bone) and 2.89%, respectively. X-ray diffraction revealed characteristic peaks of hydroxyapatite crystals (25.86°, 31°-34°). Infrared spectroscopy results showed characteristic absorption peaks for both collagen and hydroxyapatite. Thermogravimetric analysis indicated a mineral phase content of 78.29% in the material. The scaffold porosity was 91.6% ± 1.1%, close to the level of natural bone tissue. Ion release curves demonstrated sustained release behavior for both magnesium and strontium ions. The in vitro degradation rate matched the ingrowth rate of new bone tissue. Cytological experiments showed that MSHA/Col significantly promoted MC3T3-E1 cell proliferation (130% increase in activity at 72 h, P < 0.001). MSHA/Col exhibited excellent efficacy in promoting osteogenic differentiation, significantly upregulating the expression of osteogenesis-related genes and proteins (Runx2, Col-Ⅰ, Opn, Ocn) (P < 0.01). Conclusion The MSHA/Col scaffold achieves dual biomimicry of natural bone in both composition and structure, and effectively promotes osteogenic differentiation at the genetic and protein levels, breaking through the functional limitations of pure hydroxyapatite mineralized collagen. This provides a new strategy for the development of functional bone repair materials

Objective: To fabricate a hydrogel loaded with inositol hexaphosphate-zinc and preliminarily evaluate its performance in self-mineralization and osteoinduction, thereby providing a theoretical basis for the development of bone regeneration materials. Methods: The hydrogel framework (designated DF0) was formed by copolymerizing methacryloyloxyethyltrimethylammonium chloride and four-armed poly(ethylene glycol) acrylate, followed by sequentially loading inositol hexaphosphate anions via electrostatic interaction and zinc ions via chelation. The hydrogel loaded only with inositol hexaphosphate anions was named DF1, while the co-loaded hydrogel was named DF2. The self-mineralization efficacy of the DF0 , DF1 and DF2 hydrogels was characterized using scanning electron microscopy, transmission electron microscopy (TEM), energy dispersive spectroscopy (EDS), and selected area electron diffraction (SAED). The biocompatibility was assessed via live/dead cell staining and a CCK-8 assay. The osteoinductive capacity of the DF0 , DF1 and DF2 hydrogels on MC3T3-E1 cells was assessed via alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining. In the aforementioned cell experiments, cells cultured in standard medium served as the control group Results: The DF0, DF1, and DF2 hydrogels were successfully synthesized. Notably, DF1 and DF2 exhibited distinct self-mineralization within 6 days. Results from TEM, EDS, and SAED confirmed that the mineralization products were amorphous calcium phosphate in group DF1, and amorphous calciumzinc phosphate in group DF2. Biocompatibility tests revealed that none of the hydrogels (DF0, DF1, and DF2) adversely affected cell viability or proliferation. In osteogenic induction experiments, both ALP and ARS staining were intensified in the DF1 and DF2 groups, with the most profound staining observed in the DF2 group. Conclusion The developed inositol hexaphosphate-zinc hydrogel (DF2) demonstrates the dual capacity to generate calcium-phosphate compounds through self-mineralization while exhibiting excellent osteoinductive properties. This biocompatible, dual-promoting osteogenic hydrogel presents a novel strategy for bone regeneration.

ObjectiveTo analyze the clinicopathological features of patients with comorbidities of chronic hepatitis B （CHB） and steatotic liver disease （SLD）， to investigate the impact of hyperglycemia on the risk of liver fibrosis and end-stage liver diseases （ESLD）， and to provide a basis for the clinical management of such population. MethodsA total of 668 adult patients with CHB-SLD confirmed by liver biopsy in The Fifth Medical Center of Chinese PLA General Hospital from January 2011 to December 2019 were enrolled， and a retrospective cohort was established with the time of liver biopsy as the baseline and the onset of ESLD as the endpoint. All patients were followed up to March 31， 2024. Propensity score matching （PSM） was performed at a ratio of 1∶4 to balance baseline features between groups， resulting in 82 patients in the hyperglycemia group and 281 in the non-hyperglycemia group. The two groups were compared in terms of metabolic profiles， laboratory markers， and histopathological features. The Mann-Whitney U test was used for comparison of non-normally distributed quantitative data between two groups. The chi-square test or Fisher exact test was used for comparison of categorical data between two groups. A multivariate Logistic regression analysis was used to investigate the influencing factors for advanced fibrosis （AF）， and the Kaplan-Meier survival analysis and the Cox proportional-hazards regression model were used to determine the influencing factors for the development of ESLD. ResultsCompared with the non-hyperglycemia group， the hyperglycemia group had a significantly higher number of factors for metabolic disorders， a significantly higher degree of hepatic steatosis， and a significantly higher detection rate of AF （all P<0.05）. The multivariate Logistic regression analysis showed that hyperglycemia was a risk factor for AF （odds ratio = 1.753， 95% confidence interval ［CI］： 1.017　—　3.023， P=0.043）. The survival analysis showed that hyperglycemia increased the risk of ESLD （χ²=4.340， P=0.037）. The multivariate Cox regression analysis confirmed that hyperglycemia was a significant metabolic risk factor for ESLD in patients with AF （adjusted hazard ratio=3.208， 95%CI： 1.201　—　8.568， P=0.020）. ConclusionHyperglycemia can increase the risk of AF and ESLD in CHB-SLD patients. Clinical monitoring and active management should be strengthened for patients who have already developed AF and hyperglycemia.

This paper summarizes Professor LIU Fengbin's clinical experience in treating autoimmune liver disease （AILD） using the method of nourishing yin and removing stasis based on stage differentiation. He believes that the pathogenesis of AILD generally involves both deficiency in essence and excess in manifestation， with essence deficiency often presenting as liver and kidney yin deficiency， which may progress to spleen deficiency and yang deficiency over time. The excess manifestation commonly includes qi stagnation， blood stasis， damp-heat， and phlegm toxicity. Clinically， he advocates for the treatment principle of nourishing yin and removing stasis. On the foundation of nourishing liver and kidney yin， different pathological factors causing stasis are eliminated according to their nature. Treatment is also tailored to different stages of AILD. In the early and asymptomatic stages， liver qi stagnation and spleen deficiency are prominent， warranting a therapeutic approach of soothing the liver， regulating qi and strengthening the spleen. The modified Chaishao Qizhi Decoction （柴芍气滞汤） is used. During the symptomatic stage， pathogenic factors become more pronounced， often accompanied by a significant deficiency of vital qi， with damp-heat， water retention， and phlegm toxicity as key pathological features. The treatment should focus on strengthening the spleen and dispelling dampness， using modified Sijunzi Decoction （四君子汤） combined with Yinchen Wuling Powder （茵陈五苓散）. In the liver function decompensation stage， vital qi is severely deficient while pathogenic factors persist， with damp-heat， phlegm toxicity， and blood stasis obstructing the liver collaterals. Treatment should focus on nourishing blood， softening the liver， strengthening the spleen， and resolving stasis， using the modified Ruangan Yangxue Decoction （软肝养血汤）. Throughout the treatment process， emphasis is placed on tonifying the liver and kidneys while protecting yin fluids.

Objective:To analyze the latent prevalence of hepatitis B and type 2 diabetes and their correlation through an observational study.Methods:This study used a case-control design. The cases with diabetes were recruited through the diabetes management system and village doctors, while the controls without diabetes were screened from volunteers recruited by village health clinics. Capillary blood samples were collected from the study participants for the measurement of real-time blood glucose level, and venous blood samples were taken from them for the detections of HBV serological markers. Firth logistic regression model was used to fit the relationship between HBsAg positive status and diabetes status.Results:The study included 1 218 diabetes patients, 62 patients with impaired fasting glucose and 491 cases without diabetes. In the cases without diagnosis of diabetes, 11.15% had impaired fasting blood glucose and 4.43% had diabetes. Among those who reported no or unknown diagnosis of hepatitis B, 1.73% were positive for HBsAg, while 18.80% were positive for both HBV core antibody and surface antibody, indicating latent infection of hepatitis B virus. In the non-diabetes group, 0.81% reported hepatitis B history, and in the diabetes group, 2.76% reported hepatitis B history. After adjustment, the HBsAg positive rate was higher in the diabetes group ( OR=2.90, 95% CI: 1.21-6.91). Conclusions:Both diabetes and hepatitis B exhibited a high degree of latent prevalence. The HBsAg positive rate was significantly higher in those with diabetes than in those without diabetes, indicating a potential correlation. These findings highlighted the importance of strengthened screening and management of comorbidities.

Objective To construct a discharge preparation plan for patients with head and neck cancer undergo-ing tracheotomy and explore its application effect to provide a reference for improving the at-home self-care ability of tracheostomy patients with head and neck cancer.Methods The discharge preparation plan for patients with head and neck cancer undergoing tracheotomy was constructed through literature analysis and expert consultation.A total of 160 patients with tracheotomy for head and neck cancer who were hospitalized in the Department of Oto-laryngology and Head and Neck Surgery of a tertiary A general hospital in Jinan City,Shandong Province from May 2023 to February 2025 were selected by convenience sampling method as the study subjects,and were divided into an experimental group and a control group by block randomization method,with 80 cases in each group.The ex-perimental group received a discharge preparation plan for patients undergoing tracheotomy for head and neck can-cer,while the control group received routine nursing and health education.Data from admission to 1 month after discharge were collected.The levels of mindfulness awareness,self-care ability,and discharge readiness were compared between 2 groups of patients before and after intervention.Results The constructed discharge preparation plan in-cludes 5 stages(pre-contemplation,contemplation,preparation,action,and maintenance).There were no sample dropouts in either group of patients.The experimental group had higher levels of mindfulness awareness,self-care ability and discharge readiness scores than the control group,and the differences were statistically significant(P＜0.001).Conclusion The discharge preparation plan for patients with head and neck cancer undergoing tra-cheotomy is scientifically reasonable,safe,and feasible,which can effectively improve patients' discharge preparation and mindfulness level,and enhance their self-care ability.

Objective:This study aims to investigate the differences between two groups of patients treated with nucleos（t）ide analogues（NAs）：those with undetectable HBV DNA by ultrasensitive testing and those with HBV DNA below the quantification limit. The findings will provide a basis for the clinical interpretation and rational use of ultrasensitive HBV DNA results.Methods:This cross-sectional study included patients with chronic HBV infection who were treated with NAs between May 2023 and December 2023 at the outpatient clinic and inpatient department of the Hepatology Department，The Second Hospital of Shandong University. All patients had ultrasensitive HBV DNA results that were either undetectable or less than 20 IU/ml. The group with undetectable ultrasensitive HBV DNA was defined as the target not detected（TND）group，while the group with HBV DNA levels below 20 IU/ml was defined as the limit of quantitation（LOQ）group. Data on patients' sex，age，type of NAs used，routine blood tests，liver and kidney function，quantitative hepatitis B serology，ultrasensitive HBV DNA，HBV pregenomic RNA（pgRNA），abdominal ultrasound，CT or MRI imaging were collected. Differences between the two groups in terms of demographic data and HBV markers were analyzed.Results:A total of 827 patients were included in the study，with 536 patients in the TND group and 291 in the LOQ group. Compared with the TND group，the LOQ group had a younger age（ P<0.001），and higher level of ALT，AST，HBeAg positive rate，HBsAg，and HBV pgRNA（all P<0.001）. Subgroup analyses stratified by HBeAg status，cirrhosis status，and sex showed that the LOQ group had significantly higher levels of HBsAg and HBV pgRNA compared to the TND group in all subgroups（all P<0.05）. Conclusion:The LOQ group had significantly higher levels of HBsAg and HBV pgRNA compared to the TND group.

Objective:To evaluate the changes in cortical electroencephalogram (EEG) burst suppression rate (BSR) during sevoflurane anesthesia in septic mice and the correlation with cerebral blood perfusion.Methods:Forty SPF male C57BL/6J mice, aged 8-10 weeks, weighing 22-25 g, were divided into 2 groups ( n=20 each) by the random number table method: sham operation group (Sham group) and cecal ligation perforation group (CLP group). The sepsis model was established by cecal ligation and puncture in anesthetized animals. Mice in both groups inhaled 2% sevoflurane for 2 h. During sevoflurane anesthesia, BSR (30 min as an epoch) on electroencephalogram was recorded, and the cortical cerebral blood perfusion was recorded using the laser speckle flow imaging at 30, 60, 90 and 120 min of anesthesia. Results:Compared with Sham group, the cortical EEG BSR was significantly increased, and the cortical cerebral blood perfusion was decreased during sevoflurane anesthesia in CLP group ( P<0.05). Cortical EEG BSR was negatively correlated with cortical cerebral blood perfusion ( P<0.05). Conclusions:Cortical EEG BSR increases during sevoflurane anesthesia in septic mice, which may be related to decreased cortical cerebral blood perfusion.

Chinese medicine therapy for removing gallstones is one of the methods for the treatment of cholelithiasis.In the view of Professor Liu Fengbin,attacking of external pathogens,improper diet and emotional disorders contribute to the main causes of cholelithiasis,and the pathogenesis of cholelithiasis is due to qi stagnation of both liver and gallbladder,and internal obstruction of damp-heat.The occurrence of cholelithiasis is closely related to deficiency of spleen and stomach,and is correlated with the pathological factors of turbid phlegm and blood stasis.For the Chinese medicine treatment of cholelithiasis,Professor Liu follows the principle of"treatment in accordance with three categories of etiological factors"(i.e,seasons,environment and body constitution).He advocates the integration of traditional Chinese medicine and western medicine,and is good at utilizing Lingnan herbs and distinctive herbs that can dissolve stones and remove stones.The treatment for cholelithiasis is mainly through the therapies of soothing liver and alleviating depression,clearing heat and removing dampness,and normalizing gallbladder function to remove stones,and is also supplemented by the therapies of invigorating spleen and replenishing qi,regulating qi to resolve phlegm,and activiting qi movement and blood circulation.Modified Da Chaihu Decoction plus Sijin Decoction is often used as a basic formula for treating cholelithiasis,which is mainly composed of Desmodii Styracifolii Herba,Galli Gigerii Endothelium Corneum,Bupleuri Radix,Curcumae Radix,Scutellariae Radix,Aucklandiae Radix,Aurantii Fructus Immaturus stir-fried with bran,Paeoniae Radix Rubra,Linderae Radix,and Rhei Radix et Rhizoma.