TCF12 induces ferroptosis by suppressing OTUB1-mediated SLC7A11 deubiquitination to promote cisplatin sensitivity in oral squamous cell carcinoma.

Chemotherapy resistance is a major obstacle to effective cancer treatment, and promotion of ferroptosis can suppress cisplatin resistance in tumor cells. TCF12 plays a suppressive role in oral squamous cell carcinoma (OSCC), but whether it participates in the regulation of cisplatin resistance by modulating ferroptosis remains unclear. Here, we found that TCF12 expression was decreased in OSCC cells compared with normal oral cells, and it was reduced in cisplatin (DDP)-resistant OSCC cells compared with parental cells. Moreover, overexpression of TCF12 sensitized DDP-resistant cells to DDP by promoting ferroptosis. Intriguingly, silencing TCF12 reversed the promotion effect of the ferroptosis activator RSL3 on ferroptosis and DDP sensitivity, and overexpressing TCF12 antagonized the effect of the ferroptosis inhibitor liproxstatin-1 on ferroptosis and DDP resistance. Mechanically, TCF12 promoted ubiquitination of SLC7A11 and decreased SLC7A11 protein stability through transcriptional repression of OTUB1, thereby facilitating ferroptosis. Consistently, SLC7A11 overexpression neutralized the promotion effect of TCF12 on ferroptosis and DDP sensitivity. Additionally, upregulation of TCF12 hindered the growth of mouse OSCC xenografts and enhanced the DDP sensitivity of xenografts by inducing ferroptosis. In conclusion, TCF12 enhanced DDP sensitivity in OSCC cells by promoting ferroptosis, which was achieved through modulating SLC7A11 expression via transcriptional regulation of OTUB1.

[Expression and Clinical Significance of Helper T Cells 9 and Its Sytokines Interleukin 9 in Chronic Lymphocytic Leukemia].

OBJECTIVE: To investigate the expression and clinical significance of T helper cell 9 (Th9) and its cytokine interleukin 9(IL-9) in peripheral blood of patients with chronic lymphocytic leukemia(CLL). METHODS: A total of 43 newly diagnosed patients with chronic lymphocytic leukemia in the First Affiliated Hospital of Xinjiang Medical University from June 2021 to June 2022 were selected as the case group. The patients were divided into Binet A group (13 cases), Binet B group (20 cases) and Binet C group (10 cases) by Binet staging system, and 20 healthy volunteers who underwent physical examinationin in our hospital in the same period served as control group. The proportion of Th9 cells in peripheral blood was detected by flow cytometry, the expression level of Th9 specific transcription factors PU.1 and IRF4 was detected by Western blot, and the expression level of serum cytokine IL-9 was detected by ELISA. The proportion of Th9, the expression of PU.1, IRF4 and IL-9 in each group were compared, and the correlation between the proportion of Th9, IL-9 and clinicopathological indexes of CLL patients was analyzed. RESULTS: The proportion of Th9, the expression of PU.1, IRF4 and IL-9 in CLL group were significantly higher than those in control group (P<0.05), the proportion of Th9 and the expression of IL-9 in Binet B and C group were higher than those in Binet A group (P<0.05), but there was no significant difference in the proportion of Th9 cells between Binet B group and C group (P>0.05). The expression of IL-9 in Binet C group was significantly higher than that in Binet B group (P<0.05) . The proportion of Th9 cells and IL-9 were highly expression in patients with ß2 microglobulin abnormality, IGHV unmutation, P53 abnormality and hepatosplenic lymph node enlargement(P<0.05), but not related to age and sex (P>0.05). The results of Spearman correlation analysis showed that the proportion of Th9 in patients with CLL was negatively correlated with the lymphocytic account and lymphocyte proportion(rs=-0.32，rs=-0.34). The proportion of Th9 and IL-9 were positively correlated with Binet stage, Rai stage and CLL-IPI Scoring (rs=0.79，rs=0.54，rs=0.58； rs=0.72，rs=0.63，rs=0.45), but not with WBC, CD4+ T cells and CD8+T cells (P>0.05). The proportion of Th9 was positively correlated with IL-9 (rs=0.53). CONCLUSION: Th9 cells and IL-9 are abnormally highly expressed in CLL, which is related to the poor prognosis of CLL.

Personalized RNA neoantigen vaccines stimulate T cells in pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDAC) is lethal in 88% of patients1, yet harbours mutation-derived T cell neoantigens that are suitable for vaccines 2,3. Here in a phase I trial of adjuvant autogene cevumeran, an individualized neoantigen vaccine based on uridine mRNA-lipoplex nanoparticles, we synthesized mRNA neoantigen vaccines in real time from surgically resected PDAC tumours. After surgery, we sequentially administered atezolizumab (an anti-PD-L1 immunotherapy), autogene cevumeran (a maximum of 20 neoantigens per patient) and a modified version of a four-drug chemotherapy regimen (mFOLFIRINOX, comprising folinic acid, fluorouracil, irinotecan and oxaliplatin). The end points included vaccine-induced neoantigen-specific T cells by high-threshold assays, 18-month recurrence-free survival and oncologic feasibility. We treated 16 patients with atezolizumab and autogene cevumeran, then 15 patients with mFOLFIRINOX. Autogene cevumeran was administered within 3 days of benchmarked times, was tolerable and induced de novo high-magnitude neoantigen-specific T cells in 8 out of 16 patients, with half targeting more than one vaccine neoantigen. Using a new mathematical strategy to track T cell clones (CloneTrack) and functional assays, we found that vaccine-expanded T cells comprised up to 10% of all blood T cells, re-expanded with a vaccine booster and included long-lived polyfunctional neoantigen-specific effector CD8+ T cells. At 18-month median follow-up, patients with vaccine-expanded T cells (responders) had a longer median recurrence-free survival (not reached) compared with patients without vaccine-expanded T cells (non-responders; 13.4 months, P = 0.003). Differences in the immune fitness of the patients did not confound this correlation, as responders and non-responders mounted equivalent immunity to a concurrent unrelated mRNA vaccine against SARS-CoV-2. Thus, adjuvant atezolizumab, autogene cevumeran and mFOLFIRINOX induces substantial T cell activity that may correlate with delayed PDAC recurrence.

Multiinstitutional Validation Study of Cyst Fluid Protein Biomarkers in Patients With Cystic Lesions of the Pancreas.

OBJECTIVE: Prospective evaluation of 2 clinical-molecular models in patients with unknown pathology who underwent endoscopic ultrasound with fine-needle aspiration (EUS-FNA) for a cystic lesion of the pancreas. SUMMARY OF BACKGROUND DATA: Preoperative prediction of histologic subtype (mucinous vs nonmucinous) and grade of dysplasia in patients with pancreatic cystic neoplasms is challenging. Our group has previously published 2 clinical-molecular nomograms for intraductal papillary mucinous neoplasms (IPMN) that incorporated both clinical/radiographic features and cyst fluid protein markers (sFASL, CA72-4, MMP9, IL-4). METHODS: This multiinstitutional study enrolled patients who underwent EUS-FNA for a cystic lesion of the pancreas. Treatment recommendations regarding resection were based on standard clinical, radiographic, and endoscopic features. Predicted probabilities of high-risk IPMN (high-grade dysplasia/invasive cancer) were calculated using the previously developed clinical-molecular nomograms. RESULTS: Cyst fluid was obtained from 100 patients who underwent diagnostic EUS-FNA. Within this group there were 35 patients who underwent resection, and 65 were monitored radiographically. Within the group that underwent resection, 26 had low-risk IPMN or benign non-IPMN lesions, and 9 had high-risk IPMN. Within the surveillance group, no patient progressed to resection or developed cancer after a median follow-up of 12months (range: 0.5-38). Using the clinical/radiographic nomogram alone, 2 out of 9 patients with high-risk IPMN had a predicted probability >0.5. In the clinical-molecular models, 6 of 9 patients in model 1, and 6 of 9 in model 2, had scores >0.5. CONCLUSIONS: This prospective study of patients with unknown cyst pathology further demonstrates the importance of cyst fluid protein analysis in the preoperative identification of patients with high-risk IPMN. Longer follow-up is necessary to determine if this model will be useful in clinical practice.

[Significance of Tim-3 in The Imbalance of Th17/Treg in Patients with Multiple Myeloma].

OBJECTIVE: To investigate the singnificance of Tim-3 in Th17/Treg balance in patients with multiple myeloma (MM). METHODS: Fifty-six newly diagnosed MM patients and 30 healthy people were enrolled. Flow cytometry was used to detect the expression of Tim-3 on CD4+T cells, the proportion of Th17 and Treg cell, the Th17/Treg ratio, the expression of Tim-3 in Th17 and Treg cells, and the ratio of Tim-3+Th17/Tim-3+Treg cells in two groups. ELISA was used to detect the level of cytokines IL-17 and IL-10. Moreover, to analyze the relationship between Tim-3+Th17/Tim-3+Treg balance and clinical indicator. RESULTS: Compared with the control group, the expression of Tim-3 on CD4+T cells in peripheral blood of MM patients was increased (P<0.05), the ratio of Th17 cells and Th17/Treg in MM patients were higher than those in control group (P<0.05), and the ratio of Treg cells in MM patients was lower than those in control group, which the difference showed no statistically significant (P>0.05), and the level of cytokines IL-17, IL-10 and the ratio of IL-17/IL-10 in MM patients were significant higher than those in the control group (P<0.05). The expression level of Tim-3+Th17 cells and the ratio of Tim-3+Th17/Tim-3+Treg in MM patients were higher than those in control group (P<0.05), while the expression level of Tim-3+Treg cells in MM patients were lower than that in the control group (P<0.05). The ratio of Tim-3+Th17/Tim-3+Treg in MM patients were positive correlated with ISS staging, DS staging, chromosome abnormality and sFLCR (r=0.635, r=0.501, r=0.449, r=0.587). CONCLUSION: The ratios of Th17/Treg, IL-17/IL-10 and Tim-3+Th17/Tim-3+Treg increase in MM patients, among which Tim-3+Th17/Tim-3+Treg is correlated with ISS staging, DS staging, chromosomal abnormalities and sFLCR, which suggesting that Tim-3 is involved in the imbalance of Th17/Treg in MM patients.

Endothelial cell­derived CCL15 mediates the transmigration of fibrocytes through the CCL15­CCR1 axis in vitro.

Wound healing is a complex physiological process in which fibrocytes serve a vital role. However, the mechanism underlying the recruitment of fibrocytes during wound healing remains largely unknown. The present study aimed to investigate whether endothelial cells are involved in the recruitment of fibrocytes in wound healing. To mimic the in vivo angiogenic process, a co­culture system consisting of endothelial cells and fibrocytes was achieved using a permeable Transwell co­culture system. The expression of chemokines produced by endothelial cells with or without co­culture was then measured using a gene chip. Based on the dataset from chip analysis, chemokine ligand 15 (CCL15) produced by endothelial cells was identified, which likely serves a regulatory role in mediating the transmigration of fibrocytes. Overexpression of CCL15 in endothelial cells or chemokine receptor 1 (CCR1) in fibrocytes promoted the transmigration of fibrocytes, whilst silencing the expression of CCL15 in endothelial cells or that of CCR1 in fibrocytes attenuated the transmigration of fibrocytes. Results from the present study suggested that the CCL15­CCR1 axis between endothelial cells and fibrocytes serves a vital role in mediating the recruitment of fibrocytes during wound healing.

[Incidence Analysis of Monoclonal Gammopathy of Undetermined Significance in People over 40 Years of Age].

OBJECTIVE: To study the distribution of monoclonal gammopathy of undetermined significance(MGUS) in different age, sex and ethnic people over 40 years old. METHODS: Five hundred and ninety-six people(over 40 years old) examened in the Health Examination center of the First Affiliated Hospital of Xinjiang Medical University from July 2017 to September 2017 were selected. Among 596 people, male 310, female 286, Han people 488, and Uygur ethnic people 108. According to age, 596 people were divided into 3 groups, (40-59 years old group, 60-79 years old group, over 80 years old group). First, all samples were screened by capillary serum protein electrophoresis. If the suspected monoclonal bands were found in the electrophoretogram, and then the specific protein types were determined by serum immunofixation electrophoresis. RESULTS: The total incidence of MGUS in 596 screened population was 4.027%. The incidence of MGUS in 40-59 years old group, 60-69 years old group and over 80 years old group were 1.762%, 2.929% and 10% respectively, and the differences among the groups were statistically significant(P<0.05). The incidence of MGUS in male (5.806%) was significantly higher than that in female (2.097%)(χ2=5.177，P<0.05). Binary Logistic regression analysis showed that over 80 years old and male were independent risk factors for MGUS(P=0.001, OR=4.188, 95%CI: 1.814-9.673, P=0.048, OR=2.605, 95%CI: 1.009-6.725). The types of immunoglobulin in patients with MGUS were mostly IgG, IgG(66.7%) was significantly more than IgA (29.2%)(χ2=21.375，P<0.05)，and there was no significant difference in the incidence of MGUS between people with in Kappa and Lambda. CONCLUSION: The age increase and male may increase the incidence of MGUS, the IgG is the most common type of immunoglobcdin in pathogenesis of MGUS, so the early screening should be done.

Clinical application of 3D technology for tooth autotransplantation: A case report.

This report presents two cases of tooth autotransplantation using cone-beam computed tomography (CBCT), the three-dimensional (3D) simulation dental planning software and a computer-aided rapid prototyping (CARP) model. Two hopeless teeth of adult patients were replaced as their third molar teeth. Before deciding the autotransplantation, diagnostic CBCT images were acquired and imported to SimPlant software. The SimPlant dental program was used for surgical simulation prior to autotransplantation, which created 3D images of the available donor teeth and recipient site tooth and superimposed the images to display their morphological similarity. Efficient modification of the recipient socket was designed preoperatively. The CARP model of the donor tooth was prepared as a substitute for the donor tooth that would be fit into the new recipient socket during bone preparation. Autotransplantation was favourably performed in 5-6 min. Transplanted teeth healed up without clinical abnormality. The postoperative follow-up time was up to 6 years.

The m6A pathway facilitates sex determination in Drosophila.

The conserved modification N6-methyladenosine (m6A) modulates mRNA processing and activity. Here, we establish the Drosophila system to study the m6A pathway. We first apply miCLIP to map m6A across embryogenesis, characterize its m6A 'writer' complex, validate its YTH 'readers' CG6422 and YT521-B, and generate mutants in five m6A factors. While m6A factors with additional roles in splicing are lethal, m6A-specific mutants are viable but present certain developmental and behavioural defects. Notably, m6A facilitates the master female determinant Sxl, since multiple m6A components enhance female lethality in Sxl sensitized backgrounds. The m6A pathway regulates Sxl processing directly, since miCLIP data reveal Sxl as a major intronic m6A target, and female-specific Sxl splicing is compromised in multiple m6A pathway mutants. YT521-B is a dominant m6A effector for Sxl regulation, and YT521-B overexpression can induce female-specific Sxl splicing. Overall, our transcriptomic and genetic toolkit reveals in vivo biologic function for the Drosophila m6A pathway.

[mRNA Expressions of T-bet, GATA-3, ROR γt and Foxp3 in peripheral blood of patients with chronic lymphocytic leukemia in different stages].

OBJECTIVE: This study was to investigate the mRNA expression of T-bet, GATA-3, ROR γt and Foxp3 mRNA in peripheral blood of patients with chronic lymphocytic leukemia (CLL) in different stages and explore their potential role in the pathogenesis and clinical diagnosis. METHODS: A total of 46 newly diagnosed and untreated patients with CLL was chosen as patient group, including 16 patients in the stage of Binet A, 15 in the stage of Binet B, and 15 in the stage of Binet C; 20 healthy persons were selected as controls. The quantitative fluorescence PCR was adopted to detect the mRNA expression of T-bet, GATA-3, RORγt and Foxp3 in peripheral blood mononuclear cell (PBMNC). RESULTS: (1) The expression of T-bet mRNA in patient group was lower than that in normal controls (P < 0.05), while the mRNA expression of GATA-3 mRNA, ROR γt, Foxp3 in CLL patients group were higher than that in normal controls (P < 0.05), and the ratio of T-bet/GATA-3 and RORγt/Foxp3 in CLL in patient group were lower than that in normal controls(P < 0.05); (2) The later the stage, the higher the mRNA expression of GATA-3 and Foxp3. The mRNA expression of GATA-3 in stage Binet B and stage Binet C of CLL patients were higher than that in stage Binet A (P < 0.05),and the mRNA expression of Foxp3 in stage Binet C was higher than that in stage of Binet A and Binet B (P < 0.05); the later the stage, the lower the ratio of T-bet/GATA-3 and RORγt/Foxp3. The ratio of T-bet/GATA-3 in stage of Binet A CLL patients was higher than that in stage Binet C (P < 0.05) and the ratio of RORγt/Foxp3 in stage of Binet A and stage of Binet B were higher than that in stage Binet C (P < 0.05). CONCLUSION: This study found in the level of transcription factors in CLL patients that with the process of disease, the balance shifts from Th1/Th2 and Th17/Treg to Th17 and Treg, and Treg cell may play a critical immunosuppressive role in the development of CLL.

Endodontic and surgical treatment of root damage caused by orthodontic miniscrew placement.

INTRODUCTION: Miniscrews are being increasingly used for anchorage control in orthodontics. Despite the concern over root damage caused by miniscrews, there are few reports of precise clinical evaluations and appropriate management of that damage. In the case presented herein, the root damage caused by the placement of miniscrews was repaired by root canal treatment and surgical intervention. METHODS: A 44-year-old man received orthodontic treatment for intrusion of the left maxillary first molar with a miniscrew anchorage system. During that treatment, the miniscrews had fallen out and had to be reinserted more than 6 times in the same area. Two years later, the patient complained of a spontaneous pain in the maxillary left molar region. Although the patient received root canal treatment, intraoral sinus tracts could still be detected, and the patient's discomfort persisted. Periradicular surgery revealed that the persistent infection was related to root surface damage caused by orthodontic miniscrew placement. Healing was achieved by a combination of root canal treatment and surgical intervention. RESULTS: Scanning electron microscopy of the damaged distobuccal root apex revealed a mature biofilm consisting of a network of matrix that contained mostly rod-like and spherical bacteria. At a 12-month recall checkup, the patient was free of pain. A repeat periapical radiograph revealed reduction of the pretreatment radiolucent lesion. CONCLUSIONS: More careful planning of miniscrew placement is necessary to lessen the danger of root damage. Furthermore, a precise evaluation of both root and pulpal damage and careful consideration of the choice of optimal treatment modality are needed.

[Dynamic change of IL-10 and TGF-beta1 in the liver of Echinococcus multilocularis-infected mice].

OBJECTIVE: To observe the dynamic expression and function of IL-10 and TGF-beta1 in liver of BALB/c mice infected with Echinococcus multilocularis (Em). METHODS: Sixty female BALB/c mice were randomly divided into experiment group and control group. Mice in the experiment group were each injected with 0.2 ml Em protoscolex suspension (containing about 400 protoscoleces) , while those in control group received same volume of normal saline. At 2, 8, 30, 90, 180, and 360 d after infection, 5 mice from each group were sacrificed and liver specimens were collected for pathological examination and immunohistochemical detection for IL-10 and TGF-beta1. RESULTS: In mice of the experiment group, Em cysts in different sizes were found in the abdominal cavity and the liver tissue, which gradually enlarged with the time. HE staining showed infiltration of lymphocytes in liver tissue, pathological change between the cyst wall and hepatic cells. In the control, the liver lobules showed integrity and inflammatory cells were seen occasionally. The level of IL-10 expression in liver tissue of the infected mice increased with the time, and reached a peak [(1639 +/- 1.73) %] at 90 d post-infection and maintained a high level thereafter. The expression of TGF-beta1 also reached the highest level [(23.69 +/- 2.29) %] . Both were significantly higher than the control (P < 0.01), though a low level expression was found in the control at 90d post-injection. CONCLUSION: The expressions of IL-10 and TGF-beta1 both increase in the middle and late stages of the infection. Besides, their inhibited functions do not be helpful for clearing and controlling Echinococcus multilocularis infection in livers.

Implant displacement into the mandible: an unusual complication during implant surgery.

In contrast with problems regarding initial stability of dental implants in the posterior maxilla, heat generation during bone drilling is one of the complications that can occur in the mandible which is usually composed of dense bone. This report presents an unusual case of implant displacement into the mandibular posterior area of a middle-aged woman. This problem was presumably caused by poor bone density, loss of cortical bone engagement, and differences in bone quality between the alveolar bone and the basal bone.