HMGB1, a potential regulator of tumor microenvironment in KSHV-infected endothelial cells.

High-mobility group box 1 (HMGB1) is a protein that binds to DNA and participates in various cellular processes, including DNA repair, transcription, and inflammation. It is also associated with cancer progression and therapeutic resistance. Despite its known role in promoting tumor growth and immune evasion in the tumor microenvironment, the contribution of HMGB1 to the development of Kaposi's sarcoma (KS) is not well understood. We investigated the effect of HMGB1 on KS pathogenesis using immortalized human endothelial cells infected with Kaposi's sarcoma-associated human herpes virus (KSHV). Our results showed that a higher amount of HMGB1 was detected in the supernatant of KSHV-infected cells compared to that of mock-infected cells, indicating that KSHV infection induced the secretion of HMGB1 in human endothelial cells. By generating HMGB1 knockout clones from immortalized human endothelial cells using CRISPR/Cas9, we elucidated the role of HMGB1 in KSHV-infected endothelial cells. Our findings indicate that the absence of HMGB1 did not induce lytic replication in KSHV-infected cells, but the cell viability of KSHV-infected cells was decreased in both 2D and 3D cultures. Through the antibody array for cytokines and growth factors, CXCL5, PDGF-AA, G-CSF, Emmprin, IL-17A, and VEGF were found to be suppressed in HMGB1 KO KSHV-infected cells compared to the KSHV-infected wild-type control. Mechanistically, phosphorylation of p38 would be associated with transcriptional regulation of CXCL5, PDGF-A and VEGF. These observations suggest that HMGB1 may play a critical role in KS pathogenesis by regulating cytokine and growth factor secretion and emphasize its potential as a therapeutic target for KS by modulating the tumor microenvironment.

CXCL1 confers a survival advantage in Kaposi's sarcoma-associated herpesvirus-infected human endothelial cells through STAT3 phosphorylation.

Many cytokines produced by Kaposi's sarcoma-associated herpesvirus (KSHV)-infected cells have been shown to participate in the pathogenesis of KSHV. Determination of the exact role of cytokines in Kaposi's sarcoma (KS) pathogenesis is limited, however, by the difficulty to manipulate the target genes in human endothelial cells. In this study, we sought to elucidate the role of cytokines in KSHV-infected human immortalized endothelial cell line (HuARLT cells) by knockout (KO) of the corresponding target genes using the CRISPR/Cas9 system. The cytokine production profile of KSHV-infected HuARLT cells was analyzed using a protein array, and several cytokines were found to be highly upregulated following KSHV infection. This study focused on CXCL1, which was investigated by knocked out in HuARLT cells. KSHV-infected CXCL1 KO cells underwent increased cell death compared to KSHV-infected wild-type (WT) cells and mock-infected CXCL1 KO cells. Lytic replication was not observed in KSHV-infected WT nor CXCL1 KO cells. Phosphorylation of STAT3 was significantly suppressed in KSHV-infected CXCL1 KO cells. Additionally, inhibitors of STAT3 and CXCL1 induced cell death in KSHV-infected endothelial cells. Our results show that CXCL1 production is required for the survival of KSHV-infected endothelial cells, and the CXCL1 to STAT3 phosphorylation signaling pathway may be a therapeutic target for KS.

Harvesting the Entire Seventh Costal Cartilage for Secondary Rhinoplasty.

As rhinoplasty is becoming increasingly common, the number of revision cases is also increasing. These cases require more cartilage, and costal cartilage is considered a good material. We introduce a method of harvesting the entire seventh costal cartilage. Methods: This study was performed from April 2019 to March 2020. The seventh costal cartilage was harvested from 156 patients. The incision was placed at the inframammary fold in women and directly above the cartilage in men. The length of incision was approximately 3.5 cm. After skin incision, we found a muscle fascia plane, and wide dissection was performed. We opened the muscle fascia and split the rectus abdominis muscles. Then, the seventh costal cartilage was exposed. Careful subperichondrial dissection was continued circumferentially. After the posterior aspect of the cartilage was exposed, the costochondral junction was cut and dislocated. The remaining perichondral dissection was performed under direct vision to avoid violating the perichondrium. Then, we dislocated the synchondrosis between the sixth and seventh cartilages and the seventh and eighth cartilages in turn. Finally, we cut the sternocostal area while protecting the underlying perichondrium and removed the cartilage. We checked for pneumothorax and then closed the wound layer by layer using 2-0 and 3-0 Vicryl sutures. Skin closure was performed with Steri-Strips. Results: The mean time for cartilage harvesting was 52 minutes. The average length of the cartilage was 9.33 mm. No cases of pneumothorax or postoperative morbidities were noted. Conclusion: We used sufficient amounts of cartilage to perform satisfactory rhinoplasty.

Composition of Vaginal Microbiota in Pregnant Women With Aerobic Vaginitis.

Vaginal dysbiosis, such as bacterial vaginosis (BV) and aerobic vaginitis (AV), is an important cause of premature birth in pregnant women. However, there is very little research on vaginal microbial distribution in AV compared to that in BV. This study aimed to analyze the composition of the vaginal microbiota of pregnant women with AV using microbial community analysis and identify the causative organism using each criterion of the AV scoring system. Also, we compared the quantification of aerobic bacteria using quantitative polymerase chain reaction (qPCR) and their relative abundances (RA) using metagenomics. This prospective case-control study included 228 pregnant Korean women from our previous study. A wet mount test was conducted on 159 women to diagnose AV using the AV scoring system. Vaginal samples were analyzed using metagenomics, Gram staining for Nugent score determination, conventional culture, and qPCR for Staphylococcus spp., Streptococcus spp., and Enterobacteriaceae. The relative abundances (RAs) of eleven species showed significant differences among the three groups (Normal flora (NF), mild AV, and moderate AV). Three species including Lactobacillus crispatus were significantly lower in the AV groups than in the NF group, while eight species were higher in the AV groups, particularly moderate AV. The decrease in the RA of L. crispatus was common in three criteria of the AV scoring system (Lactobacillary, WBC, and background flora grades), while it did not show a significant difference among the three grade groups of the toxic leukocyte criterion. Also, the RAs of anaerobes, such as Gardnerella and Megasphaera, were higher in the AV groups, particularly moderate AV, while the RAs of aerobes were very low (RA < 0.01). Therefore, qPCR was performed for aerobes (Staphylococcus spp., Streptococcus spp., and Enterobacteriaceae); however, their quantification did not show a higher level in the AV groups when compared to that in the NF group. Therefore, AV might be affected by the RA of Lactobacillus spp. and the main anaerobes, such as Gardnerella spp. Activation of leukocytes under specific conditions might convert them to toxic leukocytes, despite high levels of L. crispatus. Thus, the pathogenesis of AV can be evaluated under such conditions.

Rab27b regulates extracellular vesicle production in cells infected with Kaposi's sarcoma-associated herpesvirus to promote cell survival and persistent infection.

Extracellular vesicles (EVs) play a crucial role in cell-to-cell communication. EVs and viruses share several properties related to their structure and the biogenesis machinery in cells. EVs from virus-infected cells play a key role in virus spread and suppression using various loading molecules, such as viral proteins, host proteins, and microRNAs. However, it remains unclear how and why viruses regulate EV production inside host cells. The purpose of this study is to investigate the molecular mechanisms underlying EV production and their roles in Kaposi's sarcoma-associated herpesvirus (KSHV)-infected cells. Here, we found that KSHV induced EV production in human endothelial cells via Rab-27b upregulation. The suppression of Rab27b expression in KSHV-infected cells enhanced cell death by increasing autophagic flux and autolysosome formation. Our results indicate that Rab27b regulates EV biogenesis to promote cell survival and persistent viral infection during KSHV infection, thereby providing novel insights into the crucial role of Rab-27b in the KSHV life cycle.

Dimethyl Sulfoxide Enhances Kaposi's Sarcoma-Associated Herpesvirus Production During Lytic Replication.

Kaposi's sarcoma-associated herpesvirus (KSHV) is an etiologic agent of Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman disease. In studies of KSHV, efficient virus production and isolation are essential. Reactivation of KSHV can be initiated by treating latently infected cells with chemicals, such as 12-O-tetradecanoyl-phorbol-13-acetate and sodium butyrate. These chemicals have been used as tools to induce lytic replication and viral production in KSHV-producing cell lines. Dimethyl sulfoxide (DMSO) is an organosulfur compound that is frequently used as an aprotic solvent similar to water. In experiments exploring signaling pathways in KSHV-infected cells, DMSO treatment alone as a vehicle affected the lytic gene expression of KSHV. However, to the best of our knowledge, the effects of DMSO on KSHV-producing cells have not yet been reported. Therefore, in this study, we investigated whether DMSO could be used as a reagent to enhance viral production during lytic replication in KSHV-producing cells and assessed the underlying mechanisms. The effects of DMSO on KSHV production were analyzed in iSLK BAC16 cells, which have been widely used for recombinant KSHV production. We found that the production of KSHV virions was significantly increased by treatment with DMSO during the induction of lytic replication. Mechanistically, lytic genes of KSHV were enhanced by DMSO treatment, which was correlated with virion production. Additionally, DMSO induced the phosphorylation of JNK during lytic replication, and inhibition of JNK abolished the effects of DMSO on lytic replication and virion production. Our findings showed that additional treatment with DMSO during the induction of lytic replication significantly improved the yield of KSHV production.

Lipopolysaccharide-Preconditioned Periodontal Ligament Stem Cells Induce M1 Polarization of Macrophages through Extracellular Vesicles.

Periodontitis is a common disease characterized by chronic inflammation and tissue destruction of gums. Human periodontal ligament stem cells (PDLSCs), derived from the periodontium, have stem cell properties similar to those of mesenchymal stem cells. PDLSCs possess not only the potential to differentiate into other tissues, but also immunomodulatory abilities. Macrophages play a critical role in periodontal disease, but little is known regarding the role of PDLSCs in macrophage modulation during inflammation. In this study, we investigated the effect of PDLSCs on the macrophage cell line. While the conditioned media from PDLSCs under normal culture conditions did not affect macrophage polarization, the lipopolysaccharide (LPS)-preconditioned PDLSCs induced significant changes in M1 polarization. Extracellular vesicles (EVs) isolated from the conditioned media of LPS-preconditioned PDLSCs induced strong M1 polarization of macrophages. Additionally, the M1 polarization was abolished by DNase I treatment of EVs. Therefore, the LPS-stimulated PDLSCs induce M1 polarization of macrophages through EVs, suggesting that the EVs from PDLSCs might be a potential therapeutic target for inflammation in the periodontium.

Asian Rhinoplasty with Rib Cartilage.

An Asian rhinoplasty is one of the most popular procedures in plastic surgery. The anatomical characteristics of the Asian nose are quite different from those of other races, including low dorsum height, short columella, a thick soft tissue covering on the tip with flaccid lower lateral cartilage, and a sunken midface with relative protrusion of the mouth due to maxilla or premaxillary retrusion. For augmentation and lengthening of the nose, a silicone implant has been commonly used in Asian countries. However, many patients suffer from silicone-related complications, which induce soft tissue contraction and deform the already fragile nasal structure. Additionally, revision surgery is also increasing in frequency due to greater patient sophistication and higher expectation that the end rhinoplasty result to be more harmonious with the patient's overall facial structure. In these circumstances, a rhinoplasty using autologous rib cartilage, giving strong support and enough amount of the cartilage to correct deformed structure and midface skeletal retrusion. If properly performed with enough experience, a rib cartilage rhinoplasty will provide excellent and long-lasting results with low risk.

Endoscopic transnasal approach for the treatment of medial orbital blow-out fracture: a technique for controlling the fractured wall with a balloon catheter and Merocel.

The authors repaired a medial blow-out fracture by using an endoscopic transnasal technique with a balloon catheter and Merocel packing in 17 subjects. The follow-up periods were from 6 weeks to 2 years, and averaged 6 months. The length of the operation was 50 minutes on average. The enophthalmos was corrected in seven of the eight patients. Supporting material for the fractured medial orbital wall was kept in place for 1 to 3 weeks. The mean volume of balloon inflation was 2 cc. The result was satisfactory. No complications resulted from the transnasal endoscopic technique. This endoscopic transnasal approach allows for a better aesthetic result because it eliminates external scarring and permits a direct approach to the medial orbital wall and has a superior visualization. A balloon catheter was used to support the fractured medial orbital fracture, which was adapted, ballooned, and then visualized using a radiopaque dye (Visipaque) in 11 cases. A postoperative computed tomographic scan revealed that this is a very useful method for controlling the status of the reduced orbital wall and eliminates the possibility of complications resulting from infection. A resected uncinate process was used as a bone graft material to repair the large defect in five cases. This method provides several advantages including a mucoperiosteal attached bone graft, working in the same operative field, and cost-effective surgical time. A transnasal endoscopic technique for medial orbital fracture is also very useful for releasing entrapment of the medial rectus muscle, because it directly pushes against the fractured wall and gives good exposure of the medial orbital wall.