ObjectiveTo investigate the incidence rate of primary liver cancer （PLC） and the progression of liver fibrosis in chronic hepatitis B （CHB） patients with low-level viremia （LLV） （HBV DNA<2 000 IU/mL but ≥20 IU/mL） after treatment adjustment， and to provide more robust evidence for clinical practice. MethodsA retrospective analysis was performed for the clinical data of LLV patients who initially received nucleos（t）ide analogue （NAs） for at least 48 weeks at the Fifth Medical Center of PLA General Hospital from August 2007 to April 2017 and subsequently underwent NAs adjustment due to LLV， and according to the virologic response after 48 weeks of treatment adjustment， the patients were divided into LLV group and complete virological response （CVR） group （HBV DNA<20 IU/mL）. The patients were followed up once every 3‍ ‍—‍ ‍6 months till the primary endpoint event of PLC or October 2024. The incidence rate of PLC and the progression of liver fibrosis were observed， and the progression of liver fibrosis was defined as an increase of ≥1 grade in fibrosis-4 （FIB-4） index. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of continuous data with skewed distribution between two groups； the chi-square test was used for comparison of categorical data between groups. The Kaplan-Meier method was used to calculate the cumulative incidence rate of PLC， and the Log-rank test was used for comparison between groups； the Cox regression analysis was used to investigate the risk factors for PLC， and the Logistic regression analysis was used to investigate the influencing factors for the progression of liver fibrosis. ResultsA total of 307 patients were enrolled， with a mean age of 50.0 years， and the male patients accounted for 80.5%. After 48 weeks of treatment with the adjusted NAs regimen， 254 patients （82.7%） achieved CVR， and 53 patients （17.3%） still had LLV. For the LLV group， the incidence rate of PLC was 30.2% and the rate of liver fibrosis progression was 22.6%， while for the CVR group， the incidence rate of PLC was only 13.4%， and the rate of liver fibrosis progression was 7.5%. The multivariate regression analyses showed that LLV was an independent risk factor for the onset of PLC （hazard ratio=2.623， 95% confidence interval ［CI］： 1.315‍ ‍—‍ ‍5.234， P=0.006） and the progression of liver fibrosis （odds ratio=3.213， 95%CI： 1.385‍ ‍—‍ ‍7.455， P=0.007）. ConclusionActive adjustment of treatment is needed immediately after the diagnosis of LLV to improve CVR， and if LLV persists after treatment adjustment， it is necessary to enhance the monitoring of liver fibrosis progression and PLC， so as to facilitate early diagnosis and treatment.

Objective To investigate the expression of essential meiotic endonuclease 1(EME1)in liver cancer tissue and its effect on the biological behavior of hepatoma cells.Methods The TCGA database was used to identify the differentially expressed genes between liver cancer tissue and paracancerous tissue.Immunohistochemistry and Western Blot were used to measure the expression abundance of EME1 in liver cancer tissue.A lentivirus was constructed by short hairpin RNA,and BEL-7404 cells were transfected with the lentivirus to interfere with the expression of the EME1 gene;the cells were divided into silencing group(shEME1 group)and control group(shCtrl group).Quantitative real-time PCR and Western Blot were used to measure the mRNA and protein expression levels of EME1;Celigo Image Cytometer and MTT assay were used to measure cell proliferation rate;flow cytometry was used to observe cell cycle;Caspase 3/7 activity was used to measure cell apoptosis.The independent-samples t-test was used for comparison between two groups.Results TCGA results showed that the mRNA expression level of EME1 in liver cancer tissue was 18.9 times that in paracancerous tissue(t=5.00,P<0.001),and the protein expression level of EME1 in liver cancer tissue was 7.0 times(based on immunohistochemistry:8.4±2.6 vs 1.2±0.4,t=7.55,P<0.001)or 2.5 times(based on Western Blot:249.0%±35.5%vs 100.0%±77.8%,t=3.02,P<0.05)that in paracancerous tissue.After lentivirus infection,compared with the shCtrl group,the shEME1 group had an mRNA expression level of EME1 reduced by 29.9%(29.9%±0.9%vs 100.0%±3.6%,t=32.82,P<0.001),a protein expression level of EME1 reduced by 35.7%(35.7%±14.9%vs 100.0%±28.9%,t=3.42,P<0.05),and a level of cell counting reduced by 45.1%(4 053±167 vs 8 988±477,t=16.91,P<0.001),as well as a level of cell activity reduced to 66.9%(0.518±0.046 vs 0.774±0.022,t=8.74,P<0.001)and a level of colony forming ability reduced to 29.0%(75±6 vs 260±9,t=28.92,P<0.001).Compared with the shCtrl group,the shEME1 group had a significant increase in the proportion of cells in G1 phase(49.9%vs 44.0%,t=8.96,P<0.001)and significant reductions in the proportion of cells in G2/M phase(15.9%vs 17.9%,t=9.13,P<0.001)and S phase(34.2%vs 38.1%,t=6.91,P<0.001),while Caspase 3/7 activity was enhanced by 1.5 times(145.8%±5.9%vs 100.0%±2.3%,t=12.50,P<0.001).Conclusion EME1 is highly expressed in liver cancer tissue,and silencing the EME1 gene can inhibit the proliferation of hepatoma cells and promote cell apoptosis.

The climate in the Lingnan area south China is characterized by high temperature and rainy days， and in terms of the eating habits， the local people are more addicted to raw， cold and savory food， all of which make Lingnan people prone to a constitution of upper heat and lower cold， and pathological manifestations of upper heat and lower cold. It is believed that the main pathogenesis of hyperthyroidism in Lingnan area is the upper heat and the lower cold， manifested as spleen yang deficiency and stomach fire excess， or kidney water depletion and heart fire hyperactivity， leading to upper heat and lower cold syndrome caused by disharmony of yin and yang and abnormal ascending and descending. Therefore， spleen cold and stomach heat and disharmony between the heart and the kidney are the main syndromes of hyperthyroidism in Lingnan area. Modified Huanglian Decoction （黄连汤） is commonly used. Additionally， for spleen cold and stomach heat syndrome， Fushen （Sclerotium Poriae Pararadicis） and Baizhu （Rhizoma Atractylodis Macrocephalae） can be added to supplement spleen and stomach， thereby treating both the root and the branch. In terms of the disharmony between the heart and the kidney syndrome， Muli （Concha Ostreae） is usually added to subdue yang and supplement yin， together with Wuweizi （Fructus Schisandrae Chinensis） to supplement kidney and calm heart and Shashen （Radix Adenophorae seu Glehniae） to nourish yin and engender liquid， thereby enriching kidney-water and moistening heart-yin. Modification of the formulas is suggested in accordance with the syndromes to achieve a better effect.

OBJECTIVE To provide references for the safe use of lorlatinib in clinical practice. METHODS The reporting odds ratio （ROR） method， Medicines and Healthcare Products Regulatory Agency comprehensive standard method （referred to as “MHRA method”） and the Bayesian confidence propagation neural network （BCPNN） method were used to detect adverse drug events （ADEs） signals of lorlatinib in the FDA Adverse Event Reporting System from the first quarter of 2019 to the fourth quarter of 2022. RESULTS & CONCLUSIONS Totally 114 overlapping ADEs signals of lorlatinib were detected by the three methods， among which there were 73 new suspicious ADEs signals which were not covered in the instruction of lorlatinib. When using loratinib in clinical practice， special attention should be paid to ADEs with a high number of cases and signals， such as various neurological diseases， psychiatric diseases， respiratory system， thoracic and mediastinal diseases； clinical manifestations included cerebral edema， cerebral infarction， pulmonary hypertension， mutism， decreased sexual desire， pleural effusion. The signals of mobile thrombophlebitis， radiation necrosis， mutism， vesicoureteral reflux not mentioned in the instructions were all strong in BCPNN detection with high specificity， to which we should pay attention in clinical application.

OBJECTIVE To mine and analyze severe cutaneous adverse reaction signals of 5 commonly used immune checkpoint inhibitors （ICIs）， and to provide reference for clinically safe use of drugs. METHODS Based on the FDA adverse events reporting system （FAERS） database，adverse drug events （ADEs） reports about severe cutaneous adverse reactions related to ipilimumab， nivolumab， pembrolizumab， atezolizumab and durvalumab were collected from listing in the United States to the fourth quarter of 2022. The ADE signals were mined and analyzed with reporting odds ratio （ROR） and Bayesian confidence propagation neural network （BCPNN）. RESULTS A total of 5 726 reports of severe cutaneous adverse reactions were collected， including 3 037 reports for nivolumab，1 465 reports for pembrolizumab， 130 reports for durvalumab， 429 reports for atezolizumab and 665 reports for ipilimumab. All 5 kinds of ICIs caused positive signals， the correlation degree of which was as follows： pembrolizumab＞atezolizumab＞nivolumab＞ipilimumab＞durvalumab. Stevens-Johnson syndrome（SJS） and toxic epidermal necrolysis （TEN） have been reported for all 5 ICIs， and the association was the strongest with pembrolizumab. CONCLUSIONS All 5 kinds of ICIs are associated with the risk of severe skin adverse reactions， and close attention should be paid to their clinical use， especially being cautious when using pembrolizumab. The combination of ICIs should be avoided as much as possible.

Objective:To investigate the in vitro inhibitory effect of methylene blue mediated photodynamic therapy (PDT) combined with berberine on Porphyromonas gingivalis (P.g). Methods:P.g was cultured until the middle to late log phase, and methylene blue was added to P.g suspension at different mass concentrations for 5 min, and a laser (wavelength 660 nm, power 140 mW/cm 2) was irradiated for 2 min to find the optimal concentration of methylene blue combined with the laser for in vitro inhibition of P.g. The effect of methylene blue mediated PDT on the in vitro inhibition of P.g and the effect of berberine on the growth curve of P.g were observed. The inhibitory effect of methylene blue mediated PDT and berberine on P.g was investigated by successive combined applications. The effect of methylene blue mediated PDT on P.g morphology was observed by scanning electron microscopy. The absorption peaks of each component were measured by ultraviolet spectrophotometer. Results:The best inhibition was achieved at a methylene blue mass concentration of 24.414 1 μg/ml under 660 nm laser excitation. The differences were statistically significant in both the methylene blue and PDT groups compared with the control group (all P<0.001). 0.05 mg/ml berberine had an inhibitory effect on the planktonic bacteria of P.g. After P.g was treated with methylene blue mediated PDT, the bacterial cell walls were crumpled into clusters. Compared with the control group, the number of colonies was reduced in the 0.05 mg/ml berberine group, and the difference was statistically significant ( P<0.01). The difference between the 0.05 mg/ml berberine + light group and the control group was not statistically significant ( P>0.05). When PDT was combined with berberine, there was a synergistic inhibitory effect on P.g. PDT followed by berberine shows a better inhibitory effect on bacteria, and the differences were statistically significant (all P<0.01). After the berberine treatment, the bacterial surface became smooth, and the length of the bacterial body increased compared with the control group. Conclusions:Methylene blue mediated PDT has an inhibitory effect on P.g. When combined with berberine, it has a synergistic inhibitory effect on P.g., and the inhibition effect is better when PDT is applied first and then berberine is applied in combination.

Objective:To carry out high-risk foot examination and grading combined with two examination methods for inpatients with type 2 diabetes, and explore the influencing factors of the occurrence and development of high-risk foot, and investigate their foot care behavior status.Methods:From July 2021 to January 2022, 409 patients with diabetes who were admitted to the Department of Endocrinology, Department of Geriatrics, Department of Cardiology, Urology Surgery and Department of Ophthalmology of Xuanwu Hospital of Capital Medical University were selected as research subjects by convenience sampling. The General Information Questionnaire, Standardized Process of At-risk Foot Screening and Stratification for Diabetic Patients, InIow's Screening for the High-Risk Diabetic Foot: A 60-Second Tool, and Foot Care Behavior Questionnaire for Diabetic Patients. Single factor analysis and Logistic regression analysis were used to explore the factors influencing the occurrence and development of high-risk foot in type 2 diabetes patients. A total of 409 questionnaires were distributed, and 392 valid questionnaires were collected, with an effective response rate of 95.8% (392/409) .Results:Among 392 patients, the detection rate of high-risk diabetic foot (HRDF) was 76.3% (299/392), and the proportion of high-risk foot grade 2 was the largest (193). Age ( OR=1.042, P<0.01) and years of hypertension ( OR=1.030, P<0.05) were independent influencing factors for the occurrence of HRDF, with statistically significant differences. Taking the high risk foot grade 3 as a reference, cerebrovascular disease [ OR=16.408, 95% CI (1.323, 203.417) ], diabetes course [ OR=1.066, 95% CI (1.008, 1.128) ], education level in middle school [ OR=0.180, 95% CI (0.056, 0.581) ], education level in primary school and below [ OR=0.126, 95% CI (0.019, 0.841) ] were independent influencing factors for the progress of high risk foot. The foot care behavior of high-risk foot patients with high risk levels was not superior to that of patients with low risk levels. Conclusions:The combination of the two screening methods can meet the examination needs of clinical and nursing medical staff for inpatients with type 2 diabetes. Emphasizing the influencing factors of HRDF occurrence and development can provide reference for early identification of high-risk foot.

Mucosal vaccines can effectively induce an immune response at the mucosal site and form the first line of defense against microbial invasion. The induced mucosal immunity includes the proliferation of effector T cells and the production of IgG and IgA antibodies, thereby effectively blocking microbial infection and transmission. However, after a long period of development, the transformation of mucosal vaccines into clinical use is still relatively slow. To date, fewer than ten mucosal vaccines have been approved. Only seven mucosal vaccines against coronavirus disease 2019 (COVID-19) are under investigation in clinical trials. A representative vaccine is the adenovirus type-5 vectored COVID-19 vaccine (Ad5-nCoV) developed by Chen and coworkers, which is currently in phase III clinical trials. The reason for the limited progress of mucosal vaccines may be the complicated mucosal barriers. Therefore, this review summarizes the characteristics of mucosal barriers and highlights strategies to overcome these barriers for effective mucosal vaccine delivery.

Chronic obstructive pulmonary disease (COPD) is an important healthcare problem worldwide. Often, glucocorticoid (GC) resistance develops during COPD treatment. As a classic hypoglycemic drug, metformin (MET) can be used as a treatment strategy for COPD due to its anti-inflammatory and antioxidant effects, but its specific mechanism of action is not known. We aimed to clarify the role of MET on COPD and cigarette smoke extract (CSE)-induced GC resistance. Through establishment of a COPD model in rats, we found that MET could improve lung function, reduce pathological injury, as well as reduce the level of inflammation and oxidative stress in COPD, and upregulate expression of nuclear factor E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), multidrug resistance protein 1 (MRP1), and histone deacetylase 2 (HDAC2). By establishing a model of GC resistance in human bronchial epithelial cells stimulated by CSE, we found that MET reduced secretion of interleukin-8, and could upregulate expression of Nrf2, HO-1, MRP1, and HDAC2. MET could also increase the inhibition of MRP1 efflux by MK571 significantly, and increase expression of HDAC2 mRNA and protein. In conclusion, MET may upregulate MRP1 expression by activating the Nrf2/HO-1 signaling pathway, and then regulate expression of HDAC2 protein to reduce GC resistance.

The radial migration of cortical pyramidal neurons (PNs) during corticogenesis is necessary for establishing a multilayered cerebral cortex. Neuronal migration defects are considered a critical etiology of neurodevelopmental disorders, including autism spectrum disorders (ASDs), schizophrenia, epilepsy, and intellectual disability (ID). TRIO is a high-risk candidate gene for ASDs and ID. However, its role in embryonic radial migration and the etiology of ASDs and ID are not fully understood. In this study, we found that the in vivo conditional knockout or in utero knockout of Trio in excitatory precursors in the neocortex caused aberrant polarity and halted the migration of late-born PNs. Further investigation of the underlying mechanism revealed that the interaction of the Trio N-terminal SH3 domain with Myosin X mediated the adherence of migrating neurons to radial glial fibers through regulating the membrane location of neuronal cadherin (N-cadherin). Also, independent or synergistic overexpression of RAC1 and RHOA showed different phenotypic recoveries of the abnormal neuronal migration by affecting the morphological transition and/or the glial fiber-dependent locomotion. Taken together, our findings clarify a novel mechanism of Trio in regulating N-cadherin cell surface expression via the interaction of Myosin X with its N-terminal SH3 domain. These results suggest the vital roles of the guanine nucleotide exchange factor 1 (GEF1) and GEF2 domains in regulating radial migration by activating their Rho GTPase effectors in both distinct and cooperative manners, which might be associated with the abnormal phenotypes in neurodevelopmental disorders.
Autism Spectrum Disorder/metabolism* ; Cell Movement/genetics* ; Humans ; Interneurons/metabolism* ; Neurodevelopmental Disorders/genetics* ; Neurons/metabolism* ; Rho Guanine Nucleotide Exchange Factors/genetics*