Objective To investigate the effect and mechanism of berberine (BBR) on the activity and proliferation of hepatoma cells. Methods Normal human hepatocytes (MIHA) and human hepatoma cells (Hep3B, HepG2) were respectively divided into control group (high glucose) and different concentration (0, 12.5, 25, 50 μmol/L) of BBR groups (glucose-free). CCK-8 and clone formation experiment were used to explore the effect of BBR on the viability and clone formation. SYTOX Green nucleic acid stain was used to determine the cell death and flow cytometry to detect the changes in reactive oxygen species (ROS) levels in hepatoma cells. The changes of Nrf2 and its related proteins after BBR incubation were measured by Western blotting. We incubated the proteasome inhibitor MG132 and the protein synthesis inhibitor cycloheximide (CHX) with BBR to study its effect on the Nrf2 protein. We quantified the expressions of Nrf2 and HO-1 in Hep3B cells cultured with BBR using Quantitative real-time PCR (qRT-PCR). We established a tumor model by subcutaneous injection of Hep3B cells into nude mice. To observe the effect of BBR on tumor growth, we injected tumor-bearing nude mice in the BBR groups and the control group with BBR or the same amount of normal saline, respectively. We evaluated the effect of BBR on Nrf2 and its related proteins in vivo with HE and immunohistochemical staining. Results The viability of Hep3B and HepG2 cells decreased after BBR incubation in a glucose-free environment (P<0.05). No appreciable changes were observed in MIHA cell viability under the same experiment condition. BBR could inhibit clone formation and promote the death of hepatoma cells in a concentration-dependent manner (P<0.05). We observed an increase in the ROS and a decrease in Nrf2, HO-1, and c-Myc with no significant change in Keap1 in hepatoma cells after BBR treatment. While total GSK3β remained unchanged, the treatment-induced decrease in P-GSK3β (Ser9) decreased but increase in P-GSK3β (Tyr216). MG132 co-treatment reversed the BBR-induced reduction of Nrf2 protein while CHX enhanced the BBR-induced reduction of Nrf2 protein in hepatoma cells. BBR treatment-induced increase in Nrf2 mRNA and a decrease in HO-1 mRNA (P<0.05). BBR could significantly inhibit the growth of Hep3B cells in vivo in nude mice. BBR treatment increased tumor tissue necrosis and suppressed the expression of Nrf2 and c-Myc but did not affect the Keap1 level. Conclusions BBR could inhibit the activity and proliferation of hepatoma cells and promote cell death. In vivo experiments showed that BBR injection could significantly inhibit the growth of tumor cells and lead to increased necrosis of tumor cells. We verified that BBR may lead to the death of hepatoma cells through the non-Keap1-dependent Nrf2/GSK3β pathway. We infer that the degradation of Nrf2 protein through the ubiquitin-proteasome system by the non-Keap1-dependent Nrf2/GSK3β pathway, thereby reducing the anti-oxidative stress ability of tumor cells and leading to the death of hepatoma cells ultimately.

Objective To investigate the effect and mechanism of mitochondrial translocation regulation of translocase of outer mitochondrial membrane 70 (Tom70) /polyribonucleotidyltransferase 1 (PNPT1) in hypoxic injury of rat myocardial cells. Methods (1) Rat H9C2 cardiomyocytes were divided into control group (treated with normoxia for 12 hours) and hypoxia group (treated with hypoxia for 12 hours). The apoptosis rate of cardiomyocytes was detected by flow cytometry, the expression of TUBA mRNA was detected by qPCR, and the expression of PNPT1 protein was detected by Western blotting. (2) H9C2 cells were infected with lentivirus vector carrying Tom70 sequence to up-regulate the expression of Tom70. The cells were then divided into NC group (lentivirus negative control group, containing lentivirus empty vector only), Tom70 over-expression group (lentivirus vector carrying Tom70 sequence), hypoxia+NC group, hypoxia+Tom70 over-expression group. Western blotting was used to detect the expression level of Tom70, PNPT1 protein in cells, and flow cytometry was used to detect the apoptosis rate of hypoxia+NC group and hypoxia+Tom70 over-expression group, the expression of TUBA mRNA in the hypoxia+NC group and hypoxia+Tom70 overexpression group were detected by qPCR, and the interaction between Tom70 and PNPT1 in the cells was detected by immunoprecipitation technique. Results (1) The results of flow cytometry showed that compared with control group, the apoptosis rate of myocardial cells in hypoxic group was significantly higher (P<0.05). The results of qPCR showed that compared with the control group, the content of TUBA mRNA in cells of hypoxia group was significantly decreased (P<0.05). Western blotting showed that compared with control group, the expression level of PNPT1 protein in mitochondria of hypoxic group was significantly decreased, while the expression level of PNPT1 protein in cytoplasm was significantly increased (P<0.05). (2) Western blotting showed that compared with control group and NC group, the expression of Tom70 in the Tom70 over-expression group was significantly increased (P<0.05); Under normal oxygen conditions, there was no significant difference in the expression of mitochondrial PNPT1 and cytoplasmic PNPT1 between NC group and Tom70 over-expression group; Under hypoxia condition, compared with hypoxia+NC group, the expression level of mitochondrial PNPT1 in hypoxia+Tom70 over-expression group was significantly higher, and the expression level of cytoplasmic PNPT1 was significantly lower (P<0.05). The results of immunoprecipitation showed that Tom70 and PNPT1 in cardiomyocytes could bind each other. The results of flow cytometry showed that compared with hypoxia+NC group, the apoptosis rate of hypoxia+Tom70 over-expression group decreased (P<0.05). The results of qPCR showed that compared with hypoxia+NC group, the content of TUBA mRNA in the hypoxia+Tom70 overexpression group increased (P<0.05). Conclusion Tom70 can alleviate hypoxic injury of rat myocardial cells by regulating the expression of PNPT1 mitochondria and reducing the degradation of apoptosis-related mRNA.

Chemotherapy is the main treatment for triple-negative breast cancer (TNBC) due to the lack of effective targeted therapy. At present, neoadjuvant chemotherapy (NAC) is one of the standard treatment strategies. However, the response of patients to NAC is different, and how to identify effective or resistant individuals before treatment is an urgent clinical problem to be solved. The research progress on the prediction of NAC response for TNBC has many aspects, including imageomics, tumor immunology and genomics. Combining different factors to build a prediction model can better improve the prediction efficiency. With the development of gene detection technology, single-cell sequencing technology can exclude the influence of non-tumor cells in tumor microenvironment, accurately determine tumor subtypes, reveal drug resistance mechanisms, and may be applied to NAC in the future to improve the accuracy of prediction. This article reviews the latest research progress in predicting the efficacy of TNBC NAC.

Tuberculosis (TB) is still a big threat to public health. TB in newborns includes congenital TB and neonatal TB. Although not common, rapid progress and high mortality rates must attract the attention of clinicians. Congenital TB is transmitted vertically from mother, mainly acquired in utero through haematogenous spread via the umbilical cord or at the time of delivery through aspiration or ingestion of infected amniotic fluid or cervicovaginal secretions. Neonatal TB is acquired after birth through exposure to a person with infectious TB. It is often difficult to distinguish congenital TB and neonatal TB. But treatment and management is the same. This paper discusses the prenatal (epidemiology, early detection and reasonable treatment of TB in pregnancy) and postpartum (evaluation and treatment of newborns with high risk factors for TB) management of tuberculosis in newborns in order to provide useful information for clinicians to identify congenital tuberculosis and neonatal tuberculosis in the early stage and take reasonable and effective treatment measures as soon as possible.

Currently, the bacillus Calmette-Guerin (BCG) is the only licensed vaccine against tuberculosis, but its protective effect is highly controversial. Therefore, it is particularly important to develop a new and effective tuberculosis vaccine. Meanwhile, with the in-depth understanding of the pathogenic mechanism of Mycobacterium tuberculosis and the continuous update of vaccine research and development technology, there will be a major breakthrough in tuberculosis vaccine. Considering the safety, economic cost and protective effect, multi-stage antigen subunit vaccine or multi-epitope vaccine has good application prospects. This paper systematically reviews the types and characteristics of current tuberculosis vaccines, and discuss the problems and challenges in four aspects: antigen selection, animal model selection, the evaluation indicators of vaccine efficacy and the population specificity in the research of new tuberculosis vaccines, which lays a theoretical foundation for the research of new tuberculosis vaccine.

Objective To observe the pathological features of diabetic tubulopathy (DT) and analyze the relationship between interstitial fibrosis and tubular atrophy (IFTA) scores and poor disease prognosis. Methods The pathological types, basic data, laboratory examination indexes and follow-up of 121 patients diagnosed as diabetic tubulopathy by pathological biopsy from 2018-2021 at the Army Specialized Medical Center of Army Medical University were collected and retrospectively analyzed. The patients were divided into 3 groups according to the different IFTA scores: group 1 (n=20), group 2 (n=55), and group 3 (n=46). Adverse prognosis was based on the occurrence of endpoint events. Differences between clinical and laboratory data of each group were compared, and correlation analysis was performed between IFTA and glomerular injury, interstitial inflammation, renal artery vitreous lesion and renal arteriosclerosis. Cox regression analysis was used to assess the relationship between IFTA scores and endpoint events. Results The age of 121 DT patients was (53.0±10.2) years old, including 84 males and 37 females, and 99 cases of the 121 patients were with hypertension (81.8%). The incidence of nocturia, lower limb edema, proteinuria, and diabetic retinopathy increased with increasing IFTA score (P<0.01), and urinary microalbumin, urinary protein quantification, urinary albumin creatinine ratio (ACR), corrected urinary N-acyl-β-D-amino glucosidase (NAG), cystatin C (CYC), and blood creatinine levels showed increasing trend (P<0.01), and urinary creatinine (Ucr) and estimated glomerular filtration rate (eGFR) levels gradually decreased (P<0.01), but there was no statistically significant difference between the development of complications such as coronary heart disease, cerebrovascular disease, and chronic liver disease and high IFTA scores (P>0.05). Among pathological changes, IFTA score was strongly correlated with glomerular injury (r=0.503, P=0.000), and renal artery vitreous changes (r=0.329, P=0.007). There was a positive correlation between IFTA score and poor prognosis of patients (model-corrected P=0.021, HR=2.740, 95%CI 1.167-6.410), where the risk of adverse prognosis was 2.740 times higher in patients with IFTA score 3 than in patients with IFTA score 1. Conclusion Glomerular injury, renal artery vitreous degeneration, and renal arteriosclerosis scores were closely associated with IFTA scores in DT, and IFTA scores were closely associated with poor prognosis in DT patients.

Obstructive sleep apnea hypopnea syndrome (OSAHS) is a common sleep disorder, which is caused by collapse and obstruction of upper airway during sleep, accompanied by snoring, daytime drowsiness and sleep structure disorder. OSAHS is closely related to cardiovascular disease, metabolic disorder, diabetes, cognitive impairment and tumors. The prevalence rate of OSAHS is relatively high, which significantly increases the social and economic burden. Therefore, early diagnosis and treatment of OSAHS is necessary. miRNA is a non-coding small RNA molecule, which can play important role in a variety of important biological functions by regulating the expression of target genes. In recent years, some studies indicated that miRNA could be used as a biomarker for early diagnosis of hypertension, coronary heart disease, diabetes and tumors. Recently, a large number of studies showed that miRNA can be used in diagnosis, assessment, and therapeutic target of OSAHS with its complications. This review summarizes the research progress on the role of miRNA in OSAHS with its complications.

Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis (MTB) that seriously affects human health. As one of the TB high-burden countries worldwide, there is a need to strengthen the prevention and treatment of tuberculosis in China. It is estimated that the proportion of people with latent TB infection (LTBI) in China is close to 20% of the total population. Effective identification of LTBI, thus enabling early diagnosis and preventive treatment, is one of the key measures to control the TB epidemic. Existing methods of diagnosing TB infection are unable to distinguish LTBI from active TB (ATB), and cannot accurately predict progression of LTBI to ATB, while biomarkers are expected to have the potential in differentiating LTBI from ATB and predict progression. In this paper, we summarize the recent advances in biomarkers associated with LTBI and address the future development trends, in order to provide ideas for finding new markers for precisely identifying LTBI.

Traumatic brain injury (TBI) is a serious public health problem, and it is estimated that more than 50 million patients worldwide su ff ered from TBI each year. At present, the diagnosis of TBI mainly relies on scoring scales, and it is extremely necessary to screen objective biomarkers of TBI. A large number of studies have focused on the screening of protein markers of TBI, however, protein markers have short half-life and low sensitivity, so it is important to screen new biomarkers of TBI for its rapid diagnostic and treatment. As a lack of effective therapeatic drugs, TBI was mainly treated according to symptoms. microRNAs (miRNAs) are a class of small non-coding RNAs whose altered expression levels are associated with a variety of diseases in central nervous system including TBI and play an important role in the regulation of neuroplasticity and repair of neuronal damage. In addition, its small molecular weight make it crossing the blood-brain barrier conveniently and be easily detected in peripheral fluids. Therefore, miRNAs have the potential to be used as diagnostic markers and therapeutic targets for TBI. This paper aims to review the features of miRNAs and their applications on diagnosis and treatment of TBI, and provide a reference for their potential clinical applications.

Objective To analyze the value of tumor growth pattern (TGP) and tumor budding (TB) on judgment of the prognosis of patients with stage Ⅰ-Ⅲ colorectal cancer. Methods The clinical data of 152 patients with stage Ⅰ-Ⅲ colorectal cancer, admitted and undergone radical operation from December 2012 to June 2020 in Hainan Hospital of Chinese PLA General Hospital, were collected and retrospectively analyzed. TGP and TB were interpreted with HE staining and patients were divided into TGP-A (dilatant growth type and intermediate type) or B (invasive growth type), TB-low or TB-high subgroups. The differences of clinicopathological features among these subgroups were analyzed; Kaplan-Meier analysis was performed to analyze the disease-free survival (DFS) and overall survival (OS) of these subgroups. Finally, Cox proportional risk model was used to analyze the risk factors influencing prognosis of colorectal cancer patients. Results Statistical differences of tumor differentiation degree and N stage existed in TGP-A or TGP-B, TB-low or TB-high subgroups (P<0.01); in addition, the characteristics were more obvious of high CEA level (P=0.03), T₃+T₄ stage (P<0.01) and TNM Ⅲ stage (P<0.01) in TGP-B groups. Between TGP-A and TGP-B subgroups, and between TB-low and TB-high subgroups, the statistical significantly difference existed in DFS (TGP: Log-rank=10.06, P<0.01; TB: Log-rank=6.62, P=0.01) and OS (TGP: Log-rank=6.53, P=0.01; TB: Log-rank=4.90, P=0.03). TGP was an independent risk factor for both DFS (HR=2.95, 95%CI 1.54-5.63, P<0.01) and OS (HR=2.37, 95%CI 1.14-4.94, P=0.02). Conclusion Both TGP and TB were more reliable prognostic markers for patients with stage Ⅰ-Ⅲ colorectal cancer, the ones presented TGP with infiltrative pattern or TB ≥10/0.785 mm² would have poor prognosis.