PI3K/AKT mediated De novo fatty acid synthesis regulates RIG-1/MDA-5-dependent type I IFN responses in BVDV-infected CD8+T cells.

Bovine viral diarrhea virus (BVDV) has caused massive economic losses in the cattle business worldwide. Fatty acid synthase (FASN), a key enzyme of the fatty acid synthesis (FAS) pathway, has been shown to support virus replication. To investigate the role of fatty acids (FAs) in BVDV infection, we infected CD8+T lymphocytes obtained from healthy cattle with BVDV in vitro. During early cytopathic (CP) and noncytopathic (NCP) BVDV infection in CD8+ T cells, there is an increase in de novo lipid biosynthesis, resulting in elevated levels of free fatty acids (FFAs) and triglycerides (TG). BVDV infection promotes de novo lipid biosynthesis in a dose-dependent manner. Treatment with the FASN inhibitor C75 significantly reduces the phosphorylation of PI3K and AKT in BVDV-infected CD8+ T cells, while inhibition of PI3K with LY294002 decreases FASN expression. Both CP and NCP BVDV strains promote de novo fatty acid synthesis by activating the PI3K/AKT pathway. Further investigation shows that pharmacological inhibitors targeting FASN and PI3K concurrently reduce FFAs, TG levels, and ATP production, effectively inhibiting BVDV replication. Conversely, the in vitro supplementation of oleic acid (OA) to replace fatty acids successfully restored BVDV replication, underscoring the impact of abnormal de novo fatty acid metabolism on BVDV replication. Intriguingly, during BVDV infection of CD8+T cells, the use of FASN inhibitors prompted the production of IFN-α and IFN-ß, as well as the expression of interferon-stimulated genes (ISGs). Moreover, FASN inhibitors induce TBK-1 phosphorylation through the activation of RIG-1 and MDA-5, subsequently activating IRF-3 and ultimately enhancing the IFN-1 response. In conclusion, our study demonstrates that BVDV infection activates the PI3K/AKT pathway to boost de novo fatty acid synthesis, and inhibition of FASN suppresses BVDV replication by activating the RIG-1/MDA-5-dependent IFN response.

Efficacy and safety of CalliSpheres® microspheres drug-eluting beads transarterial chemoembolization in GCLM combined trans-arterial infusion therapy for treating primary focus of gastric cancer: a multi-center retrospective study.

BACKGROUND: The purpose of this study was to observe the safety and efficacy of CalliSpheres microspheres drug-eluting beads (DEB) transhepatic arterial chemoembolization (CSM-TACE) for liver metastasis of gastric cancer (GCLM) combined with trans-arterial infusion therapy (TAIT) as the primary focus of gastric cancer. RESEARCH DESIGN AND METHODS: Unresectable advanced GCLM patients were collected for retrospective analysis. Patients who progressed after chemotherapy or could not receive systematic chemotherapy were selected. CSM-TACE was used for GCLM treatment and oxaliplatin for TAIT of primary focus of gastric cancer. Adverse reactions, tumor reactions, survival rate, and survival time during treatment were recorded, and prognostic factors were analyzed. RESULTS: Forty-three patients from four oncology centers met inclusion criteria and were enrolled. CSM-TACE averaged (1.51 ± 0.51) times and TAIT averaged (4.58 ± 1.65) times. The follow-up time was 2.5-49 months, and the 6-month, 1-year, and 2-year survival rates were 86.0%, 72.1%, and 41.9%, respectively, with a median overall survival of 11.5 months. The adverse reactions during treatment were grade 1-3. The hazard ratio (HR) of combination therapy was 0.51 (P = 0.040), and the HR of TAIT frequency was 0.34 (P = 0.002), which were independent protective factors affecting prognosis. CONCLUSIONS: CSM-TAC for GCLM combined with TAIT for primary focus of gastric cancer is safe and efficacious, which is worthy of clinical promotion and application.

Diagnostic accuracy of de-escalated surgical procedure in axilla for node-positive breast cancer patients treated with neoadjuvant systemic therapy: A systematic review and meta-analysis.

BACKGROUND: More initial clinical node-positive breast cancer patients achieve axillary pathological complete response (ax-pCR) after neoadjuvant systemic therapy (NST). Restaging axillary status and performing de-escalated surgical procedures to replace routine axillary lymph nodes dissection (ALND) is urgently needed. Targeted axillary lymph node biopsy (TLNB) is a novel de-escalated surgical strategy marking metastatic axillary nodes before NST and targeted dissection and biopsy intraoperatively to tailor individual axillary management. METHODS: This study provided a systematic review and meta-analysis to evaluate the feasibility and diagnosis accuracy of TLNB. Prospective and retrospective clinical trials on TLNB were searched from Pubmed, Embase, and Cochrane. Identification rate (IFR), false-negative rate (FNR), negative predictive value (NPV), and rate of ax-pCR were the outcomes of this meta-analysis. RESULTS: One thousand nine hundred and twenty patients attempted TLNB, with an overall IFR of 93.5% (95% confidence interval [CI] 90.1%-96.2%). IFR of three nodal marking methods, namely iodine seeds, clips, and carbon dye, was 95.6% (95% CI 91.2%-98.7%), 91.7% (95% CI 87.3%-95.4%), and 97.1% (95% CI 89.1%-100.0%), respectively. Of them, 847 patients received ALND, with an overall FNR of 5.5% (95% CI 3.3%-8.0%), and NPV ranged from 90.1% to 96.1%. Regression analysis showed that the overlap of targeted and sentinel biopsied nodes might associate with IFRs and FNRs. CONCLUSION: TLNB is a novel, less invasive surgical approach to distinguish initial node-positive breast cancer that achieves negative axillary conversion after NST. It yields an excellent IFR with a low FNR and a high NPV. A combination of preoperative imaging, intraoperative TLNB with SLNB, and postoperative nodal radiotherapy might affect the future treatment paradigm of primary breast cancer with nodal metastases.

Roles and mechanisms of miR-195-5p in human solid cancers.

Cancer persists as a worldwide disease that contributes to high morbidity and mortality rates. As a class of non-coding RNA, microRNAs (miRNAs) are one kind of important regulators in cancer and frequently implicated in tumor development and progression. Emerging experiments have suggested that miRNA-195-5p (miR-195-5p) can regulate neoplastic processes in many pathways. For instance, miR-195-5p can not only regulate proliferation, migration and invasion of tumor cells but also promote tumor cell apoptosis. Furthermore, low expression of miR-195-5p could induce drug resistance. Our review focuses on the expression of miR-195-5p in various tumors and elucidates the related mechanisms of which miR-195-5p participates in tumor biology, as well as summarizes the roles of miR-195-5p in tumor progression. We believe that miR-195-5p might have potential utility as a novel diagnostic biomarker and therapeutic target for cancer.

[Accuracy and safety of robot assisted pedicle screw placement].

OBJECTIVE: To investigate the accuracy and safety of pedicle screw placement assisted by orthopedic robot and C-arm fluoroscopy. METHODS: The clinical data of 36 patients with spinal diseases underwent surgical treatment from January 2019 to August 2020 was retrospectively analyzed. Among them, 18 cases were implanted pedicle screws assisted by orthopaedic robot(observation group), including 12 males and 6 females, aged from 16 to 61 years with an average of (38.44±3.60) years;there were 1 case of adolescent scoliosis, 1 case of spinal tuberculosis, 7 cases of lumbar spondylolisthesis, 4 cases of thoracic fracture and 5 cases of lumbar fracture. Another 18 cases were implanted pedicle screws assisted by C-arm fluoroscopy(control group), including 10 males and 8 females, aged from 18 to 58 years with an average of (43.22±2.53) years;there were 1 case of adolescent scoliosis, 6 cases of lumbar spondylolisthesis, 6 cases of thoracic fracture and 5 cases of lumbar fracture. The intraoperative fluoroscopy times, nail placement time and postoperative complications were recorded in two groups. CT scan was performed after operation. The Gertzbein-Robbins standard was used to evaluate the accuracy of pedicle screw placement which was calculated. RESULTS: The number of intraoperative fluoroscopy in observation group was(6.89±0.20) times, which was significantly higher than that in control group(14.00±0.18)times(P<0.05). The placement time of each screw in observation group was(2.56±0.12) min, which was significantly different from that in control group(4.22±0.17) min (P<0.05). One case of incision infection occurred in control group after operation, and recovered after active dressing change. During the follow-up period, no serious complications such as screw loosening and fracture occurred in two groups, and there was no significant difference in complications between two groups(P>0.05). A total of 107 screws were placed in observation group, including 101 screws in class A, 4 in class B, 2 in class C, 0 in class D and 0 in class E, the accuracy rate of pedicle screw placement=[(number of screws in class A+B) / the number of all screws placed in the group] ×100%=98.1%(105/107); and a total of 104 screws were placed in control group, including 90 screws in class A, 4 in class B, 5 in class C, 5 in class D and 0 in class E, the accuracy rate of pedicle screw implantation=[(number of screws in class A+B/the number of all screws placed in the group]×100%=90.3% (94/104); there was significant difference between two groups (P<0.05). CONCLUSION: Orthopaedic robot assisted pedicle screw placement has the advantages of less fluoroscopy times, shorter screw placement time and higher accuracy, which can further improve the surgical safety and has a broad application prospect in the orthopaedic.

Screening of the novel immune-suppressive biomarkers of TMED family and whether knockdown of TMED2/3/4/9 inhibits cell migration and invasion in breast cancer.

Background: Transmembrane p24 trafficking protein (TMED) family members are implicated in several solid tumors, but their clinical relevance for breast cancer (BC) remains unclear. This study aimed to probe their prognostic values and relations with tumor immunity in BC. Methods: TMED family mRNA expression was assessed in five microarray datasets (GSE65212, GSE42568, GSE5364, GSE22820 and GSE45827) from Gene Expression Omnibus (GEO) database and invasive breast cancer (BRCA) cohort from The Cancer Genome Atlas (TCGA). Receiver operating characteristic (ROC) curve was performed to determine the predictive values of filtered members of the TMED family. The protein expressions of screen genes were validated by Clinical Proteomic Tumor Analysis Consortium (CPTAC) data from University of ALabama at Birmingham CANcer data analysis portal (UALCAN) and detected in the clinical specimens by western blot assay. Clinicopathologic variables were analyzed with bc-GenExMiner, and patient prognostic data were obtained with Kaplan-Meier Plotter. In vitro wound healing and invasion assays were performed on siRNA-transfected BC cell lines. TIMER 2.0, SangerBox, and ImmPort were used to evaluate tumor immune infiltration, immune checkpoints, and other immune-related genes. CbioPortal, Metascape, Expression2kinases, and LinkedOmics were used to explore gene regulatory network. Results: BC tissues expressed TMED2/3/4/9 at a higher level than normal tissues, providing diagnostic potential. All the areas under the ROC curve for TMED2/3/4/9 were more than 0.7. TMED2/3/4/9 correlated with numerous clinical variables, including lymph node status, Scarff-Bloom-Richardson score (SBR), Nottingham Prognostic Index (NPI), estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER-2), and triple-negative breast cancer (TNBC) status, and their high expression predicted the poor prognosis of BC patients. TMED2/3/4/9 knockdown drastically inhibited the migratory and invasive capacities of MDA-MB-231 and HCC1937 cells. TMED2/3/4/9 expressions correlated negatively with the infiltration of tumor-suppressive immune cells such as CD8+ T cells, dendritic cells, and natural killer cells, and was inversely related to a variety of immune checkpoint genes, including programmed cell death 1 (PD-1) and cytotoxic T-lymphocyte associated protein 4 (CTLA4). A set of kinases, transcription factors, and microRNAs (miRNAs) may regulate TMED2/3/4/9 abnormalities at the genome level. Conclusions: TMED2/3/4/9 may serve as diagnostic, prognostic, and immune-suppressive biomarkers in BC.

[Clinical study of modified one stage posterior approach total en block spondylectomy and spinal reconstruction in the treatment of invasive thoracic vascular tumor].

OBJECTIVE: To explore the surgical method and safety of modified one stage posterior approach total en block spondylectomy combined with pedicle screw fixation and titanium mesh reconstruction for the treatment of invasive thoracic vascular tumor. METHODS: The clinical data of 12 patients with invasive thoracic vasculay tumor from December 2012 to May 2015 was retrospectively analyzed. There were 8 males and 4 females, aged from 40 to 62 years with an average of 51.2 years, the course of disease was 2 months to 8 years with an average of 3.4 years. The lesions involved vertebral bodies:1 case of T2, 4 cases of T4, 1 case of T5, 2 cases of T6, 2 cases of T8, and 2 cases of T10. According to Tomita classification, there were 3 cases of typeⅠ, 3 cases of typeⅡ, 1 case of type Ⅲ, and 5 cases of type Ⅳ. The Japanese Orthopaedic Association (JOA) score was 8.0±2.7 before operation. One patient with T10 lesions had sensory dyskinesia below the umbilicus, and the muscle strength of both lower limbs was grade Ⅰ-Ⅱ, and the others 11 patients were grade Ⅲ-Ⅳ. All 12 patients underwent one stage posterior approach total en block spondylectomy and the pedicle screw fixation combined with titanium mesh reconstruction under general anesthesia and continuous motor evoked potential (MEP) spinal cord electrophysiological monitoring throughout the operation. The operation time, intraoperative blood loss and transfusion, postoperative pain and recovery of spinal cord function, bone graft fusion, tumor recurrence and other complications were followed up. RESULTS: All the operations were successful. The average operation time, intraoperative blood loss and blood transfusion were 5.5 h (4.5 to 6.0 h), 1 850 ml (1 650 to 2 500 ml), 1 050 ml (600 to 1 500 ml), respectively. All 12 patients were followed up for 5 months to 2.5 years with an average of 21 months. Local pain and lower limb muscle strength were improved to varying, and the nerve compression symptoms disappeared. The JOA score at 6 months after operation was 12.0±3.4, which was statistically significant difference compared with the preoperative 8.0±2.7 (t=3.20, P<0.05). Titanium mesh bone grafts were all fused in phaseⅠ, with an average fusion time of 4.5 months (3 to 7 months). During the follow-up period, there was no tumor recurrence, loosening or breaking of nails, sinking and displacement of titanium mesh. CONCLUSION: Modified one stage posterior approach total en block spondylectomy is an ideal surgical method for the treatment of invasive thoracic vascular tumors, which has a safe, reliable and long lasting efficacy.

[Treatment of degenerative lumbar spondylolisthesis with minimally invasive transforaminal lumbar interbody fusion by microscope-assisted Zista channel].

OBJECTIVE: To investigate the clinical efficacy and advantage of minimally invasive transforaminal lumbar interbody fusion under microscope-assisted Zista channel in the treatment of degreeⅠandⅡdegenerative lumbar spondylolisthesis. METHODS: The clinical data of 18 patients with degenerative lumbar spondylolisthesis treated by microscope-assisted Zista channel MIS-TLIF operation from January 2017 to March 2018 were analyzed retrospectively. There were 10 males and 8 females with an average age of 59 years (48 to 70). The course of spondylolisthesis ranged from 6 months to 5 years with an average of 33 months. The segment of spondylolisthesis was L4 in 13 casesand L5 in 5 cases. According to Meyerding spondylolisthesis criteria, 11 cases were degreeⅠand 7 cases were degreeⅡ. The perioperative observation indexes were recorded. The VAS score and JOA score of lumbago and leg pain before and after operation were compared to evaluate the clinical efficacy. At 12 months after operation, the height of intervertebral space was measured by imaging data, the intervertebral fusion was evaluated by Brantigan-Steffee standard, and the curative effect was evaluated according to the modified Macnab standard. RESULTS: All the patients completed the operation successfully and were followed up more than 12 months after operation. Operation time was(160.45±34.98) min, intraoperative blood loss was (88.32±21.12) ml, postoperative drainage volume was (50.34 ±18.22)ml, and walking time after operation was (20.65±6.25) h. Preoperative and postoperative at 7 days, 3 months, 12 months, VAS score of low back pain was 7.81±2.16, 4.19±1.17, 2.25±0.62 and 1.53±0.58 respectively, VAS score of leg pain was 8.47± 2.21, 3.45±0.86, 2.31±0.73 and 1.43±0.47, JOA score was 12.01±2.33, 18.56±3.12, 23.54±3.31 and 26.34±2.65. There were significant differences in VAS and JOA scores between preoperative and postoperative (P<0.05). The height of intervertebral space increased from (4.46±0.72) mm preoperative to (10.24±1.48) mm at 12 months after operation (P<0.05). All operative segments got fusion (16 cases of grade E and 2 cases of grade D). According to Macnab standard, 15 cases obtained excellent results, 2 cases good, 1 case fair. CONCLUSION: MIS-TLIF under microscope-assisted Zista channel has obvious minimally invasive advantages in the treatment of degreeⅠandⅡdegenerative lumbar spondylolisthesis, and it is a safe and effective method.

Dark tea extracts: Chemical constituents and modulatory effect on gastrointestinal function.

Processing of dark tea varieties, such as Fu brick tea, Liupao tea, Qianliang tea, and Qing brick tea, includes solid-state fermentation involving microorganisms. In this study, we analyzed the major chemical constituents of dark tea extracts and evaluated their modulatory effect on the gastrointestinal function in normal mice, including the improvement of gastrointestinal transit and intestinal microbial, as well as the attenuation of intestinal microbial dysbiosis and intestinal pathological damage, and the adjustment of immune function in antibiotic-treated mice. Substantial differences in major chemical constituents, including total polyphenols, total organic acids, water extract content, 18 free amino acids, gallic acid, and six tea catechins, were observed among Fu brick tea, Qianliang tea, Qing brick tea, and Liupao tea extracts. Extracts from the four dark tea varieties significantly promoted gastrointestinal transit and colonization of beneficial Bifidobacterium and Lactobacillus, and inhibited the growth of harmful Escherichia coli and Enterococcus in normal mice. In addition, Qianliang tea, Qing brick tea, and Liupao tea extracts significantly accelerated the reversal of the ampicillin sodium-induced pathological damage in the ileum, intestinal bacterial dysbiosis (Bifidobacterium, Lactobacillus, E. coli, and Enterococcus), and low immunity.

[Application of total en bloc spondylectomy in spinal tumors].

Total en bloc spondylectomy is a surgical technique cutting off tumors of spine and its satellite. Previous studies have shown that this technique could reduce postoperative recurrence rate of spinal tumor, but surgical indications are not unified. Wide application of spinal tumor surgical staging and life expectancy system make it more clear for surgery. However, it is difficult to carry out extensive operation for the characteristics of more bleeding, great difficulty and high risk. As for continuous improvement of TES technology, application of endoscopy and appearance of 3D printing artificial vertebral body could push the operation becoming mature.

[Retrograde interlocked intramedullary nailing with tibia bone graft fusion for the treatment of end-stage ankle arthritis].

OBJECTIVE: To explore clinical outcomes of retrograde interlocked intramedullary nailing with tibia bone graft fusion in treating end-stage ankle arthritis. METHODS: From November 2014 to April 2016, 22 patients with end-stage ankle arthritis were treated with retrograde interlocked intramedullary nailing with tibia bone graft fusion, including 9 males and 13 females aged from 30 to 65 years old with an average of 48.5 years old. Seven patients had obvious varus deformity, and other 15 patients' appearance were normal. Operative time, blood loss, fracture healing time and complications were observed and compared, AOFAS and VAS score were applied for evaluate ankle joint function and pain degree before operation and 9 months after operation. RESULTS: All patients were followed up from 12 to 24 months with an average of 18.6 months. The incision were healed at stage I , and no complications occurred. Operative time ranged from 80 to 120 min with an average of 90 min;blood loss ranged from 15 to 50 ml with an average of 30 ml;fracture healing time was from 10 to 18 weeks with an average of 14 weeks. AOFAS score at 9 months after operation was 88.00±3.45, while 54.82±2.98 before operation, and there was statistical significance; 8 cases obtained excellent results, 12 good and 2 moderate. There was significant difference in VAS score between preoperative(3.96±1.27) and 9 months after operation(9.37±0.55). CONCLUSIONS: Retrograde interlocked intramedullary nailing with tibia bone graft fusion for the treatment of end-stage ankle arthritis has advantages of stable fixation, less trauma, less blood loss, bone union and rapid recovery of function, and could relieve pain obviously.

[Progress on research of Madelung's deformity].

Madelung deformity is a rare deformity of forearm and wrist caused by growth disorders of distal radius ulnar and palmar epiphyseal. Current studies showed that its incidence mainly associated with trauma, epiphyseal developmental abnormalities, nutritional disorders and genetic deletion or mutation. The early clinical presentation is not typical, in middle and late time, wrist deformity and weak can appear. Plain film considered as the main means of diagnosis is often lack of early diagnosis significance. Although wrist joint magnetic resonance imaging showing early soft tissue and skeletal abnormalities were used for the early diagnosis of the disease, current domestic study in magnetic resonance imaging of this deformity is less. According to the size of the distal ulnar inclination angle and palm angle, this deformity can be divided into different types. The patients with severe deformity and symptoms usually need surgical intervention including ulna revision and osteotomy of the distal radius at present. Although the two operation can achieve good clinical results, the surgical trauma, infection and postoperative risk of joint activities are more.