Dihydroartemisinin inhibits HNSCC invasion and migration by controlling miR-195-5p expression.

Objectives: Dihydroartemisinin (DHA), an artemisinin derivative extracted from the traditional Chinese medicinal herb Artemisia annua, has the potential to suppress head and neck squamous cell carcinoma (HNSCC) progression. However, the mechanisms underlying these effects remain unclear. Therefore, we aimed to examine the mechanisms underlying the effects of DHA on tumor invasion and migration. Methods: Human HNSCC cell lines CAL-27 and FaDu were exposed to varying DHA concentrations (0, 5, 20, and 80 µM) for 24 h. Cell proliferation, invasion, and migration were assessed using CCK8, transwell, and wound-healing assays, respectively. Quantitative real-time PCR, western blotting, and immunofluorescence were used to assess the expression levels of the target genes and proteins. Results: DHA suppressed the invasion and migration of CAL-27 and FaDu cells. Additionally, miR-195-5p suppressed the invasion and migration of HNSCC cells. This study revealed significant differences in the expression of miR-195-5p and TENM2 between clinical samples and multiple public databases. DHA treatment and miR-195-5p overexpression significantly reduced TENM2 expression in HNSCC cells, which suggested that miR-195-5p overexpression enhanced the inhibitory effect of DHA on TENM2. Conclusions: This study provides the first evidence that DHA inhibits cell invasion and migration by regulating the miR-195-5p/TENM2 axis in HNSCC cells, suggesting it as a potentially effective treatment strategy for HNSCC.

A novel cuproptosis-related gene signature of prognosis and immune microenvironment in head and neck squamous cell carcinoma cancer.

BACKGROUND: Cuproptosis is a novel form of cell death that is highly related to mitochondrial metabolism and mediated by protein lipoacetylation. This study systematically assessed the differential expression and genetic alterations of cuproptosis-related genes (CRGs) in head and neck squamous-cell carcinoma (HNSCC) and constructed CRG risk models to predict survival in patients with HNSCC. METHODS: We investigated the expression of 19 CRGs in HNSCC and noncancerous tissues, and the relationship between mutation load, immune infiltration, and clinical features was examined based on data from public databases. CRG risk models were constructed by univariate Cox analysis and lasso regression and validated by independent datasets for their accuracy in predicting survival outcomes in patients with HNSCC. The expression distribution of CRGs in HNSCC cells was further explored in the HNSCC single-cell sequencing dataset. RESULTS: NFE2L2, ATP7A, FDX1, LIAS, DLD, DLAT, PDHB, MTF1 and DBT were highly expressed in noncancerous samples, while GLS, CDKN2A and DLST were highly expressed in HNSCC samples (p < 0.05). Gene copy number variation frequency (CNV) revealed CDKN2A, FDX1 and DLAT copy number deletions and LIPT2 and NFE2L2 copy number increases. Ten CRGs were used to construct a risk model to predict overall survival (OS) in HNSCC, yielding reduced OS in the high-risk group compared to the low-risk group, training group (p = 9.733e - 05), and testing group (p = 0.040). The CRG risk model was significantly correlated with various immune cells, regulatory T cells (Tregs) and memory B cells were significantly negatively correlated (p = 0.027, p = 0.00084), and resting CD4 memory T cells was significantly positively correlated (p = 9e - 04). Most CRGs significantly affected the clinical characteristics of HNSCC. NFE2L2, SLC31A1, PDHA1, CDKN2A and DBT were highly expressed in epithelial cells of HNSCC, while SLC31A1, DBT and NFE2L2 were highly expressed in T cells, and SLC31A1 in B cells. In monocytes, NFE2L2, SLC31A1 and PDHA1 were highly expressed. CONCLUSION: The CRG risk model can be used as a potential prognostic factor for HNSCC patients and may provide new insights into cancer treatment from the perspective of copper metabolism.

Icaritin inhibits PD-L1 expression by Targeting Protein IκB Kinase α.

Icaritin, a small molecule currently being investigated in phase III clinical trials in China (NCT03236636 and NCT03236649) for treatment of advanced hepatocellular carcinoma (HCC), is a prenylflavonoid derivative obtained from the Epimedium genus. Previously, it was found that Icaritin decreased the expression of PD-L1, but its direct molecular targets and the underlying mechanisms have not been identified. In this study, we report the identification of IKK-α as the protein target of Icaritin by biotin-based affinity binding assay. The further mutagenesis assay has provided evidence that C46 and C178 in IKK-α were essential amino acids for Icaritin binding to IKK-α, revealing the binding sites of Icaritin to IKK-α for the first time. Functionally, Icaritin inhibited the NF-κB signalling pathway by blocking IKK complex formation, which led to decreased nuclear translocation of NF-κB p65, and subsequent downregulation of PD-L1 expression in a dose-dependent manner. More importantly, PD-L1-positive patients exhibited longer overall survival upon Icaritin therapy. Finally, Icaritin in combination with checkpoints antibodies, such as α-PD-1, has demonstrated much better efficacy than any single therapy in animal models. This is the first report that anticancer effects of Icaritin are mediated, at least in part, by impairing functions of IKK-α.

[Clinical investigate of functional exercise immediate after operation for Achilles tendon rupture].

OBJECTIVE: To investigate clinical effect of partly weight-bearing walking and functional exercise immediatly after operation for Achilles tendon rupture(ATR) on function of ankle joint and rate of fragmentation of Achilles tendon, through comparing effect of partly weight-bearing walking and functional exercise immediatly at 2 weeks after operation for Achilles tendon rupture. METHODS: Sixty-four patients with ATR selected from March 2012 to March 2013 were randomly divided into two groups. There were 34 patients in treatment group, including 18 males and 16 females with an average age of 41.4±7.6, they began to do functional exercise and walk on fields with partly weight-bearing at two days after operation; there were 30 patients in control group, including 16 males and 14 females with an average age of 39.9±7.6, and they were immobilized with plaster in plantar flexion at two weeks after operation, and started to do functional exercise and walk on fields with partly weight-bearing at two weeks after operation. Two groups were performed by the same doctor with the same operation. The rate of fragmentation of Achilles tendon, and AOFAS score and complications between two groups were observed and compared. RESULTS: AOFAS score in treatment group at two weeks after operation was 74.3±3.9, which in control group was 71.7±4.2, and had statistical differences between two groups; AOFAS score in treatment group at one year after operation was 93.3±3.9, which in control group was 92.0±4.1, and had no statistical significance. No Achilles tendon fragmentation in treatment group occurred at three years after operation, and 1 patient occurred in control group. Two patients in treatment group occurred complications after operation, and 1 patient occurred in control group, however, there was no statistical significance between two groups. CONCLUSIONS: Functional exercise immediate after operation for Achilles tendon rupture(ATR) patients in the early days, the AOFAS scores is higher than the fixing for two weeks, and does not increase the rate of fragmentation of Achilles tendon and complication after operation, and benefits for function recovery.

IL-8 Secreted from M2 Macrophages Promoted Prostate Tumorigenesis via STAT3/MALAT1 Pathway.

Prostate cancer (PCa) is a major health problem in males. Metastasis-associated with lung adenocarcinoma transcript-1 (MALAT1), which is overexpressed in PCa tissue, is associated with physiological and pathological conditions of PCa. M2 macrophages are major immune cells abundant in the tumor microenvironment. However, it remains unknown whether M2 macrophages are involved in the effects or not, and molecular mechanisms of MALAT1 on PCa progression have not yet been comprehensively explored. Here we reported that, M2 macrophages (PMA/IL-4 treated THP1) induced MALAT1 expression in PCa cell lines. Knockdown MALAT1 expression level in PCa cell lines inhibited cellular proliferation, invasion, and tumor formation. Further mechanistic dissection revealed that M2 macrophages secreted IL-8 was sufficient to drive up MALAT1 expression level via activating STAT3 signaling pathway. Additional chromatin immunoprecipitation (ChIP) and luciferase reporter assays displayed that STAT3 could bind to the MALAT1 promoter region and transcriptionally stimulate the MALAT1 expression. In summary, our present study identified the IL-8/STAT3/MALAT1 axis as key regulators during prostate tumorigenesis and therefore demonstrated a new mechanism for the MALAT1 transcriptional regulation.

Clinical observation of biomimetic mineralized collagen artificial bone putty for bone reconstruction of calcaneus fracture.

This study investigated clinical outcomes of biomimetic mineralized collagen artificial bone putty for bone reconstruction in the treatment of calcaneus fracture. Sixty cases of calcaneal fractures surgically treated with open reduction and internal fixation in our hospital from June 2014-2015 were chosen and randomly divided into two groups, including 30 cases treated with biomimetic mineralized collagen artificial bone putty as treatment group, and 30 cases treated with autogenous ilia as control group. The average follow-up time was 17.2 ± 3.0 months. The results showed that the surgery duration and postoperative drainage volume of treatment group were significantly lower than control group; there were no statistically significant differences in the fracture healing time, American Orthopaedic Foot and Ankle Society scores at 3 and 12 months after surgery, Böhler's angle, Gissane's angle and height of calcaneus between the two groups. There were no significant differences in wound complication and reject reaction between the two groups, while significant difference in donor site complication. As a conclusion, the implantation of biomimetic mineralized collagen artificial bone putty in the open reduction of calcaneal fracture resulted in reliable effect and less complications, which is suitable for clinical applications in the treatment of bone defect in calcaneal fractures.

[The surgical options of cervical tuberculous lymphadenitis].

OBJECTIVE: To analyze the clinical features & variation features of cervical tuberculous lymphadenitis and to discuss its effective surgical treatment. METHOD: Retrospective analysis of 27 cases of cervical tuberculous lymphadenitis patients admitted to the hospital from January 2008 to December 2013. The cervical tuberculous lymphadenitis is classified based on the enhanced CT scanning conducted before operation and the preoperative evaluation of clinical characters of patients. The lymphaden ncisional biopsy was conducted on the type I patients with cervical tuberculous lymphadenitis. The lesion resection was performed on the type II and mixed type I + II patients. The regional cervical lymph node dissection was carried on the type III, the type IV and other mixed type patients. The negative pressure drainage ball was placed after operation, and the cavity was flushed with 5% povidone iodine solution. The antituberculosis therapy was performed after wound healing. RESULT: The analysis of the clinical features for 27 patients: the incidence rate on the left side, right side, both sides and middle-line is 63.0%, 25.9%, 7.4% and 3.7% respectively. The majority of patients whose lesion involving more than one region account for 62.1%; the patients whose lesion involving one region account for 37.9%. The most common is level V involved lesion (69.0%), then level IV (62.1%), level III (51.7%), level II (34.5%), level I (10.3%) and level VI (3.4%) in order. The analysis of the CT imageology features for 27 patients: the simple type is the majority (65.5%), and the most common is type III (24.1%), then the type I (17.2%), type II (13.8%) and type IV (10.3%) in order. The mixed type is minority (34.5%), but the two mixed is often (31.0%). The three mixed is only located on one side (3.4%). The recurrence never happened on the patients with resection and standard antituberculosis therapy. CONCLUSION: For the cervical tuberculous lymphadenitis, the suitable surgical treatments shall be selected according to the lesion characteristics & location and CT imaging manifestations presurgical evaluation. The effective way to treat cervical tuberculous lymphadenitis is to conduct negative pressure drainage after operation, to flush the cavity with 5% povidone iodine solution and to perform antituberculosis therapy.

[A case of the removing the intragastric foreign body by the gastroscopy combining with the esophagoscopy].

A male patient, 49 years old with drug use, swallowed a toothbrush and a tube of 20 centimeters long which lead to the upper abdomen pain for six years. The gastroscopy examination showed the head of the toothbrush in the gastric antrum and the tail inserting in the pylorus. The intragastric toothbrush was removed by the gastroscopy combining with esophagoscopy.

Radiolocalization of sentinel lymph nodes in clinically N0 laryngeal and hypopharyngeal cancers.

OBJECTIVES: We sought to analyze the characteristics of radioactive lymph nodes with metastatic disease and to explore methods for the localization of sentinel lymph nodes (SLNs) with radionuclide in clinically N0 laryngeal and hypopharyngeal cancer. METHODS: Forty-five patients with T1-T4 and clinically N0 laryngeal and hypopharyngeal cancer were recruited. For each patient, a peritumoral submucosal injection of 99mTc-labeled sulfur colloid was administered, and lymph node mapping was performed by lymphoscintigraphy 2 hours after injection. The SLNs were localized during operation by a hand-held gamma probe 10 to 12 hours after the injection, and we defined the radioactive counts from the parotideomasseteric region as background values. All lymph nodes that had accumulated radioactivity were harvested and initially termed as SLNs. Selective neck dissection was performed in all patients. The SLN specimens were sent for formal paraffin-embedded sectioning, serial sectioning, and immunohistochemical assay. The results were compared to those for the remaining lymphadenectomy specimen. Resection of the primary tumor depended on its location and the T classification. RESULTS: Sentinel lymph nodes were identified in 41 of 45 patients (51 necks). Sentinel lymph nodes with occult metastases were found in 13 patients (15 necks). In a false-negative case, metastasis was found in a nonsentinel lymph node in 1 of the neck specimens. The SLN identification rate was 92.7%, the sensitivity was 93.7%, the false-negative rate was 6.3%, and the accuracy was 98.0%. In 11 of the 15 necks (73.3%) with pathologically positive SLNs, metastasis was found in the node with the highest radioactivity. Harvesting the first 3 nodes with the highest radioactive counts may identify patients with occult metastatic disease. CONCLUSIONS: Excision of the first 3 SLNs with the highest radioactive counts can be used to accurately identify the status of cervical lymph node metastases in patients with clinically N0 laryngeal or hypopharyngeal cancer.

[Localization of sentinel lymph node with radionuclide in clinically N0 laryngeal and hypopharyngeal cancers].

OBJECTIVE: To analyze the characteristic of the radioactive lymph node with metastatic disease and to explore the method of the localization of sentinel lymph node (SLN) with radionuclide in N0 clinically laryngeal and hypopharyngeal cancer. METHODS: Fourty-five patients with T1-T4 and clinically N0 laryngeal and hypopharyngeal cancer were recruited. For each patient a peritumoral submucosal injection of 99mTc-labeled sulfur colloid (99mTc-SC) was performed and lymph node mapping was performed by lymphoscintigraphy two hours afterward. The SLN was localized by a handheld gamma probe intraoperatively 10-12 hours after the injection. All hot lymph nodes accumulating activity were harvested and initially termed sentinel nodes. Selective neck dissections were performed for all patients. The specimen of SLN was sent to the pathologist for the following analysis: formal paraffin embedded section, consecutive section and immunohistochemistry assay. The results was compared to the remaining lymphadenectomy specimen. Resection of the primary tumour depended on the location and the T classification. RESULTS: SLNs were identified in 41 of 45 patients with 51 necks, SLNs had occult metastases in 13 cases, 15 necks with SLN-positive of these 13 cases, there was one false negative case, they were found in non-SLNs of neck specimens. Each neck side was considered a single case. SLN identification rate was 92.7%, sensitivity was 93.7%, false-negative rate was 6.3%, and accuracy was 98.0%. In 11 (73.3%) of these SLN-positive necks, the SLN with the highest counts contained tumor; harvesting the first-three nodes with the highest radioactive counts, which could all patients with occult metastatic disease. CONCLUSIONS: Excision of the first-three SLNs with the highest radioactive counts can accurately judge the presence or absence of the cervical lymph nodes metastases in patients with the clinically N0 laryngeal and hypopharyngeal cancer.