OBJECTIVES: To investigate the inhibitory effect of multimerin-2 (MMRN2) overexpression on growth and metastasis of cutaneous melanoma cells. METHODS: Clinical data of patients with cutaneous melanoma were obtained from the GEO database to compare MMRN2 expressions between normal and tumor tissues. A protein-protein interaction network was constructed using the STRING database, and the intersecting genes from GEPIA2.0 were subjected to GO and KEGG enrichment analysis. The prognostic relevance of MMRN2 expression level was assessed using Cox regression and "timeROC". The correlations of MMRN2 expression level with immune infiltration and angiogenesis-related genes were analyzed using GSCA database and the ssGSEA algorithm. Colony-forming assay, Transwell assay, and wound healing assay were used to examine the changes in proliferation and migration of cultured cutaneous melanoma cells following MMRN2 knockdown. In a mouse model bearing cutaneous melanoma xenograft, the effect of MMRN2 knockdown on vital organ pathologies, survival of the mice and GM-CSF, CXCL9, and TGF‑β1 protein expressions were analyzed. RESULTS: MMRN2 was significantly upregulated in metastatic cutaneous melanoma (P<0.001). Protein interaction network analysis identified 15 intersecting genes, which were enriched in endothelium development and cell-cell junctions. In patients with cutaneous melanoma, a high MMRN2 expression was correlated with a poor prognosis, an advanced T stage, a greater Breslow depth, and ulceration (P<0.05). MMRN2 expression level was strongly correlated with 24 immune cell types (P<0.001), fibroblasts, endothelial cells, and expressions of the pro-angiogenic genes (KCNJ8, SLCO2A1, NRP1, and COL3A1; P<0.001). In cultured B16F10 cells, MMRN2 knockdown significantly suppressed cell proliferation, migration and invasion and caused remo-deling of the immunosuppressive microenvironment. CONCLUSIONS MMRN2 overexpression drives progression of cutaneous melanoma by enhancing tumor metastasis, angiogenesis and immune evasion, highlighting its potential as a therapeutic target for melanomas.
Humans ; Melanoma/metabolism* ; Animals ; Cell Movement ; Prognosis ; Skin Neoplasms/metabolism* ; Mice ; Cell Proliferation ; Neoplasm Invasiveness ; Cell Line, Tumor ; Protein Interaction Maps

ObjectiveTo observe the effect of Shufeng Jiedu capsule （SFJD）-containing serum on human lung epithelial cells infected by influenza A virus， and investigate the protective effect of the drug on the cells and the potential antiviral effect. MethodThe SFJD-containing serum was prepared and used to treat human lung epithelial cells （BEAS-2B） cultured in vitro. The viability of cells treated with different concentrations of SFJD-containing serum was measured by the cell counting kit-8 （CCK-8）， and the optimal concentration of SFJD-containing serum was screened for subsequent experiments. BEAS-2B cells were classified into normal control， virus infection， and SFJD-containing serum groups， and the CCK-8 method was used to detect the survival rate of BEAS-2B cells after virus infection and drug administration. The expression of influenza virus nucleic acid in the cells of each group was determined， and the apoptosis of cells in different groups was observed by fluorescence microscopy. Real-time PCR was employed to determine the mRNA levels of influenza virus nucleoprotein （NP）， Toll-like receptor 4 （TLR4）， and myeloid differentiation primary response gene 88 （MyD88） in each group of cells. The immunofluorescence assay was used to detect the fluorescence intensities of TLR4， MyD88， and phosphorylated nuclear factor-κB （p-NF-κB） in lung epithelial cells. ResultCompared with that in the control group （normal serum）， the cell survival rates in the blank serum and the SFJD-containing serum （5%， 10%， and 20%） groups were 100.00%±0.00%， 89.05%±4.80%， 87.13%±5.90%， 93.83%±6.03%， and 99.33%±3.39%， respectively （P<0.01）. The SFJD-containing serum of 20% was selected as the optimal treatment for subsequent experiments. Compared with the normal control group， the virus infection group showed reduced cell survival rate （P<0.01）， and the reduction was increased by the SFJD-containing serum （P<0.01）. Compared with the virus infection group， SFJD-containing serum reduced the virus load （P<0.01） to decrease apoptosis. Compared with the normal control group， the virus infection group showed up-regulated mRNA levels of NP， TLR4， and MyD88 （P<0.01）， and the up-regulation was down-regulated by the SFJD-containing serum （P<0.05， P<0.01）. The fluorescence intensities of TLR4， MyD88， and p-NF-κB proteins in the cells increased after virus infection compared with those in the normal control （P<0.05， P<0.01）， and they were decreased after administration with the SFJD-containing serum （P<0.05）. ConclusionThe SFJD-containing serum can inhibit influenza virus in vitro by increasing the survival rate， reducing the apoptosis， and down-regulating the protein levels of TLR4， MyD88， and p-NF-κB in BEAS-2B cells.

ObjectiveTo evaluate the effectiveness of Lutongning granules in the treatment of trigeminal neuralgia in animal models and study its mechanism of action， so as to provide laboratory data support for the clinical application of Lutongning granules and precise treatment. MethodMale SD rats were randomly divided into normal group， model group， carbamazepine group （0.06 g·kg^-1·d^-1）， high-dose Lutongning group （2.70 g·kg^-1·d^-1）， and low-dose Lutongning group （1.35 g·kg^-1·d^-1） according to the stratified basic mechanical pain thresholds， with 10 rats in each group. A trigeminal neuralgia model of rats was prepared by injecting 30% talc suspension into the infraorbital foramen area of the rat. The drug groups were administered 10 mL·kg^-1 of drugs by gavage after 2 h of modeling. The normal group and the model group were administered distilled water by gavage under the same conditions once a day for 10 consecutive days. Von Frey brushes were used to determine the mechanical pain threshold of rats. A fully automated blood and body fluid analyzer was employed to detect the blood routine of rats. Hematoxylin and eosin （HE） staining was utilized to detect the pathological changes in the trigeminal ganglion and medulla oblongata tissue. Transmission electron microscopy was used to scan the ultrastructure of the medulla oblongata tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of inflammatory factors interleukin （IL）-1， IL-6， IL-8， tumor necrosis factor （TNF）-α， neuropeptide substance P， and β-endorphins （β-EP） in the serum of rats， and Western blot was used to detect the protein expression levels of IL-1β， purinergic receptor P2X7 （P2X7R）， and phosphorylated p38 mitogen-activated protein kinase （p-p38 MAPK）. ResultCompared with that in the normal group， the pain threshold of rats in the model group was significantly lower （P<0.01）. The absolute value of neutrophils （NEUT#） and the percentage of neutrophils （NEUT） were significantly improved， and the percentage of lymphocytes （LYMPH） was significantly reduced （P<0.01）. The serum levels of IL-1， IL-6， IL-8， and TNF-α were significantly increased （P<0.01）. SP content in brain tissue was significantly increased， and β-EP content was significantly decreased （P<0.01）. The relative protein expression of IL-1β， P2X7R， and p-p38 MAPK was significantly increased （P<0.05）. HE staining and transmission electron microscopy results of medulla oblongata tissue revealed neuronal degeneration， mild proliferation of microglial cells， reduction in the number of myelinated nerves， and obvious demyelination. The trigeminal nerve fibers of rats were disarranged， and some nerve fibers showed vacuolization. Axons were swollen， and Schwann cells proliferated. Demyelination was observed. Compared with the model group， each administration group significantly increased the pain threshold of rats （P<0.05， P<0.01）， reduced NEUT# and NEUT， and elevated LYMPH （P<0.05， P<0.01）. The administration group significantly decreased the levels of IL-1， IL-6， IL-8， and TNF-α in serum and SP in brain tissue （P<0.01） and increased the level of β-EP （P<0.01）. They significantly down-regulated the protein expression of IL-1β， P2X7R， and p-p38 MAPK（P<0.05， P<0.01） and significantly ameliorated the pathological changes in medulla oblongata tissue and trigeminal nerves of rats. ConclusionLutongning Granules had significant therapeutic effects on trigeminal neuralgia induced by injection of talc into the infraorbital foramen of model rats， and the mechanism may be related to amelioration of P2X7R-mediated neuroinflammation and inhibition of demyelination of myelinated nerves.

Tumor metastasis is closely related to high mortality rate of cancer.It is well known that glutamine plays an important role in the malignant progression of cancer.Notably,as an important carbon and nitrogen donor,glutamine has been found to be closely related to tumor metastasis in recent years.Glutamine is not only involved in regulating the proliferation of tumor cells,but is also closely related to the migration and invasion of tumor cells.Furthermore,various enzymes along with transporters in the metabolism of glutamine are involved in the process of tumor metastasis through different signaling pathways.This review provided a summary of the role of glutamine in tumor metastasis in recent years and proposed therapeutic targets to provide new strategies for the clinical treatment of tumor metastases.

Objective To investigate the effect of the aqueous extract of Chuan Xiong Rhizoma(CR)on brain metastasis of melanoma B16F10 cells in mice.Methods C57BL/6J mouse models of brain metastasis of melanoma were established by ultrasound-guided intraventricular injection of Luc-labeled B16F10 cells,and brain tumor growth was monitored by in vivo imaging.The mouse models were then randomized for daily gavage of saline or aqueous extract of CR(equivalent crude drug concentration of 1 mg/g).Behavioral tests were used to evaluate the neuroprotective effects of CR in the tumor-bearing mice,and the changes in proteins associated with blood-brain barrier integrity,neuronal cell proliferation and apoptosis,and microglial cell apoptosis and activation were observed using immunofluorescence assay.The efficacy of CR combined with temozolomide(25 mg/kg)against brain metastases of B16F10 cells was observed by in vivo imaging.Results CR-treated mouse models did not show obvious progression of brain metastases and had a reduced rate of body weight loss and lowered protein expressions of ZO-1,claudin-5,occludin,P-gp,TNF-α,AQP4 and PDGFRβ.In the behavioral tests,the CR-treated mice showed prolonged stay on the wooden stick with a shortened time of sticky stick removal.Immunofluorescence assay showed increased proliferation and decreased apoptosis of neuronal cells and microglia in CR-treated mice.CR treatment significantly increased the levels of CD86,CD206,IL-4 and IL-10 and decreased the levels of CD163 and IL-1β in the microenvironment of brain metastases.The mice receiving combined treatments with CR and temozolomide showed significantly lower intensity of fluorescent signals in the brain than those treated with temozolomide alone.Conclusion CR does not promote brain metastasis of melanoma while inducing opening of the blood-brain barrier,and its combined use with TMZ results in enhanced inhibition against brain metastasis of melanoma B16F10 cells in mice.

Objective:To investigate the effect and influencing factors of non-invasive correction of infants with congenital auricular deformity.Methods:The data of infants with congenital auricular deformity were retrospectively analyzed, who were treated with domestic external ear orthosis in the Department of Otolaryngology Head and Neck Surgery of Tianjin Hospital from January 1, 2019 to June 30, 2023. Before treatment, the skin was prepared routinely. First, the orthosis base was installed and pressed to fit the skin around the ear. Then, according to the type of auricle deformity, the ear hook and ear cover were installed, and the adhesive tape was fixed. During the corrective treatment, the orthosis was worn all day, and the treatment was ended if there was no obvious rebound after 1 month of observation. The complications were closely observed during treatment. After the treatment, the treatment effect was evaluated and divided into markedly effective, effective and ineffective. The markedly effective rate (markedly effective ears/total ears × 100%) and effective rate [(markedly effective ears + effective ears)/ total ears × 100%] were calculated. At the same time, the cure time of the markedly effective children was recorded, and they were grouped according to different types of auricle deformities and the initial age of correction (<2, 2-6, >6 weeks groups), and the related indicators were compared. SPSS 23.0 software was used for statistical analysis. The measurement data were expressed by Mean±SD. The comparison between multiple groups was analyzed by one-way ANOVA. The pairwise comparison between groups was performed by LSD method. The counting data was expressed by frequency (% or ratio). The comparison between multiple groups was performed by χ 2 test, and the pairwise comparison between groups was performed by Fisher test with adjusted α value. Results:A total of 31 cases (55 ears) were included, including 13 males (23 ears) and 18 females (32 ears), aged 4 to 127 d. There were 13 lop ears, 6 constricted ears, 6 cup ears, 4 Stahl’s ears, 5 prominent ears, 18 helical rim deformity ears, 2 conchal crus ears and 1 Ⅱ degree microtia ear. During the treatment, 5 ears had skin lesions, 5 ears had mild allergic reactions, and 2 ears had severe allergic reactions, who improved significantly and continued treatment after removal of the external orthosis and use of erythromycin ointment. After noninvasive correction treatment, the auricle deformity of the children was improved to different extent. The overall markedly effective rate was 70.91% (39/55), and the overall effective rate was 90.91% (50/55). The markedly effective rate of lop ear, constricted ear, cup ear, Stahl’s ear, prominent ear, helical rim deformity ear, conchal ear and Ⅱ degree microtia ear were 13/13, 4/6, 4/6, 4/4, 4/5, 8/18, 2/2, 0/1 respectively. The average cure time of 39 markedly effective ears was (36.08±14.77) d. The cure time of different auricle deformity types was statistically significant ( P<0.01). Pairwise comparison between the groups showed that the cure time of lop ears was shorter than constricted ears, cup ears, Stahl’s ear, prominent ears, and that the cure time of prominent ears was longer than lop ears, constricted ears, cup ears, Stahl’s ears, helical rim deformity ears, conchal crus ears (all P<0.05). According to different initial ages of correction, the children were divided into <2-week-old group (12 cases, 21 ears), 2-6-week-old group (10 cases, 18 ears), and >6-week-old group (9 cases, 15 ears), excluding 1 Ⅱ degree microtia ear. The markedly effective rates were (90.5%) 19/21, 13/18, and 7/15, respectively, with statistically significant differences ( P<0.05). The comparison between groups showed that the markedly effective rate of <2-week-old group was significantly higher than that of >6-week-old group ( P<0.017), but there was no statistically significant difference in the cure time among the three groups ( P>0.05). Conclusion:The effect of non-invasive correction for congenital auricular deformity in infants is significant. The type of congenital auricular deformity is a key factor that affects the therapeutic effect and cure time. In lop ear, Stahl’s ear, prominent ear and conchal crus deformity, good therapeutic effects can be seen, but in constricted ear, cup ear, helical rim deformity ear and Ⅱ degree microtia, the effects are poor. The cure time of lop ear is short, while the cure time of prominent ear is long. The better effect can be acquired in the earlier non-invasive correction.

Objective:To investigate the effect and influencing factors of non-invasive correction of infants with congenital auricular deformity.Methods:The data of infants with congenital auricular deformity were retrospectively analyzed, who were treated with domestic external ear orthosis in the Department of Otolaryngology Head and Neck Surgery of Tianjin Hospital from January 1, 2019 to June 30, 2023. Before treatment, the skin was prepared routinely. First, the orthosis base was installed and pressed to fit the skin around the ear. Then, according to the type of auricle deformity, the ear hook and ear cover were installed, and the adhesive tape was fixed. During the corrective treatment, the orthosis was worn all day, and the treatment was ended if there was no obvious rebound after 1 month of observation. The complications were closely observed during treatment. After the treatment, the treatment effect was evaluated and divided into markedly effective, effective and ineffective. The markedly effective rate (markedly effective ears/total ears × 100%) and effective rate [(markedly effective ears + effective ears)/ total ears × 100%] were calculated. At the same time, the cure time of the markedly effective children was recorded, and they were grouped according to different types of auricle deformities and the initial age of correction (<2, 2-6, >6 weeks groups), and the related indicators were compared. SPSS 23.0 software was used for statistical analysis. The measurement data were expressed by Mean±SD. The comparison between multiple groups was analyzed by one-way ANOVA. The pairwise comparison between groups was performed by LSD method. The counting data was expressed by frequency (% or ratio). The comparison between multiple groups was performed by χ 2 test, and the pairwise comparison between groups was performed by Fisher test with adjusted α value. Results:A total of 31 cases (55 ears) were included, including 13 males (23 ears) and 18 females (32 ears), aged 4 to 127 d. There were 13 lop ears, 6 constricted ears, 6 cup ears, 4 Stahl’s ears, 5 prominent ears, 18 helical rim deformity ears, 2 conchal crus ears and 1 Ⅱ degree microtia ear. During the treatment, 5 ears had skin lesions, 5 ears had mild allergic reactions, and 2 ears had severe allergic reactions, who improved significantly and continued treatment after removal of the external orthosis and use of erythromycin ointment. After noninvasive correction treatment, the auricle deformity of the children was improved to different extent. The overall markedly effective rate was 70.91% (39/55), and the overall effective rate was 90.91% (50/55). The markedly effective rate of lop ear, constricted ear, cup ear, Stahl’s ear, prominent ear, helical rim deformity ear, conchal ear and Ⅱ degree microtia ear were 13/13, 4/6, 4/6, 4/4, 4/5, 8/18, 2/2, 0/1 respectively. The average cure time of 39 markedly effective ears was (36.08±14.77) d. The cure time of different auricle deformity types was statistically significant ( P<0.01). Pairwise comparison between the groups showed that the cure time of lop ears was shorter than constricted ears, cup ears, Stahl’s ear, prominent ears, and that the cure time of prominent ears was longer than lop ears, constricted ears, cup ears, Stahl’s ears, helical rim deformity ears, conchal crus ears (all P<0.05). According to different initial ages of correction, the children were divided into <2-week-old group (12 cases, 21 ears), 2-6-week-old group (10 cases, 18 ears), and >6-week-old group (9 cases, 15 ears), excluding 1 Ⅱ degree microtia ear. The markedly effective rates were (90.5%) 19/21, 13/18, and 7/15, respectively, with statistically significant differences ( P<0.05). The comparison between groups showed that the markedly effective rate of <2-week-old group was significantly higher than that of >6-week-old group ( P<0.017), but there was no statistically significant difference in the cure time among the three groups ( P>0.05). Conclusion:The effect of non-invasive correction for congenital auricular deformity in infants is significant. The type of congenital auricular deformity is a key factor that affects the therapeutic effect and cure time. In lop ear, Stahl’s ear, prominent ear and conchal crus deformity, good therapeutic effects can be seen, but in constricted ear, cup ear, helical rim deformity ear and Ⅱ degree microtia, the effects are poor. The cure time of lop ear is short, while the cure time of prominent ear is long. The better effect can be acquired in the earlier non-invasive correction.

Objective To investigate the effect of the aqueous extract of Chuan Xiong Rhizoma(CR)on brain metastasis of melanoma B16F10 cells in mice.Methods C57BL/6J mouse models of brain metastasis of melanoma were established by ultrasound-guided intraventricular injection of Luc-labeled B16F10 cells,and brain tumor growth was monitored by in vivo imaging.The mouse models were then randomized for daily gavage of saline or aqueous extract of CR(equivalent crude drug concentration of 1 mg/g).Behavioral tests were used to evaluate the neuroprotective effects of CR in the tumor-bearing mice,and the changes in proteins associated with blood-brain barrier integrity,neuronal cell proliferation and apoptosis,and microglial cell apoptosis and activation were observed using immunofluorescence assay.The efficacy of CR combined with temozolomide(25 mg/kg)against brain metastases of B16F10 cells was observed by in vivo imaging.Results CR-treated mouse models did not show obvious progression of brain metastases and had a reduced rate of body weight loss and lowered protein expressions of ZO-1,claudin-5,occludin,P-gp,TNF-α,AQP4 and PDGFRβ.In the behavioral tests,the CR-treated mice showed prolonged stay on the wooden stick with a shortened time of sticky stick removal.Immunofluorescence assay showed increased proliferation and decreased apoptosis of neuronal cells and microglia in CR-treated mice.CR treatment significantly increased the levels of CD86,CD206,IL-4 and IL-10 and decreased the levels of CD163 and IL-1β in the microenvironment of brain metastases.The mice receiving combined treatments with CR and temozolomide showed significantly lower intensity of fluorescent signals in the brain than those treated with temozolomide alone.Conclusion CR does not promote brain metastasis of melanoma while inducing opening of the blood-brain barrier,and its combined use with TMZ results in enhanced inhibition against brain metastasis of melanoma B16F10 cells in mice.

Objective:To explore the effects of Circ_0000291 on proliferation,apoptosis and immune escape profiling of he-patocellular carcinoma(HCC)cells as a molecular sponge to regulate miR-326 through targeting polypyrimidine tract-binding protein 1(PTBP1).Methods:The expressions of Circ_0000291,miR-326 and PTBP1 in tissue samples and cells were measured by qRT-PCR.Dual luciferase reporter assay was employed to confirm the targeting relationship between Circ_0000291 and miR-326 as well as miR-326 and PTBP1.CCK-8 assay,Transwell assay,flow cytometry assay and ELISA were respectively utilized to determine the prolifera-tion,invasion,apoptosis and the expressions of immune regulation related factors(TNF-α,IFN-γ).Results:Compared with normal tissue and hepatic cells,Circ_0000291 and PTBP1 expressions were overexpressed in HCC tissue and cells,but the expression of miR-326 was obviously down-regulated(all P<0.05).Moreover,functional assays indicated that downregulation of Circ_0000291 could inhibit the proliferation,invasion,and immune escape while induce apoptosis of HCC cells(all P<0.05),which was according-ly exhibited in miR-326 transfection group.Overexpression of Circ_0000291 partly attenuated the anti-tumor effects induced by miR-326(all P<0.05).In HCC,Circ_0000291 could promote the expression of PTBP1 as a molecular sponge to regulate miR-326.Conclu-sion:Circ_0000291 served as a molecular sponge of miR-326 to increase PTBP1 expression and promote the proliferation,invasion and immune escape but inhibit apoptosis of HCC.There upon then,it facilitates the progression of HCC.

Tuberculosis (TB) remains a significant global health challenge, ranking second only to COVID-19 as the leading cause of death from a single infectious agent, with 1.3 million TB-related deaths reported in 2022. Treatment efficacy has been compromised by the emergence of drug-resistant strains, including rifampin-resistant TB (RR-TB), multidrug-resistant TB (MDR-TB), and extensively drug-resistant TB (XDR-TB). Although first-line drugs like isoniazid, rifampicin, pyrazinamide, and ethambutol form the cornerstone of TB therapy, the rise of resistant strains necessitates the use of second-line drugs, which often come with increased toxicity and limited accessibility. Recent advances have focused on repurposing existing compounds and developing new drugs with novel mechanisms of action. Promising agents such as second-generation bedaquiline analogs (TBAJ-587, TBAJ-876), sudapyridine (WX-081), delamanid, pretomanid, and TBI-166 (pyrifazimine) have shown efficacy against resistant Mtb strains. Innovative treatment regimens like the BPaLM protocol-combining bedaquiline, pretomanid, linezolid, and moxifloxacin-offer shorter, all-oral therapies with higher cure rates. Personalized treatment durations and dose optimizations are becoming feasible through risk stratification algorithms and pharmacokinetic/pharmacodynamic studies. Immunotherapy is emerging as a complementary strategy to enhance the host's immune response against Mtb. Agents such as vitamin D, corticosteroids, non-steroidal anti-inflammatory drugs (NSAIDs), statins, metformin, and biological agents like interleukins and granulocyte-macrophage colony-stimulating factor are under exploration. Additionally, cell therapies involving mesenchymal stem cells and immune effector cells present new therapeutic avenues. Despite these advancements, significant challenges remain in achieving the World Health Organization's "End TB Strategy" goals, particularly as the COVID-19 pandemic has diverted resources and attention. Ongoing research and global collaboration are crucial to develop novel therapeutic strategies, optimize treatment regimens, and ultimately reduce the global burden of TB.
Humans ; Antitubercular Agents/therapeutic use* ; COVID-19/epidemiology* ; Tuberculosis/epidemiology*