Identification and validation of a novel signature based on cell-cell communication in head and neck squamous cell carcinoma by integrated analysis of single-cell transcriptome and bulk RNA-sequencing.

Background: The heterogeneous crosstalk between tumor cells and other cells in their microenvironment means a notable difference in clinical outcomes of head and neck squamous cell carcinoma (HNSCC). CD8+ T cells and macrophages are effector factors of the immune system, which have direct killing and phagocytosis effects on tumor cells. How the evolution of their role in the tumor microenvironment influences patients clinically remains a mystery. This study aims to investigate the complex communication networks in the HNSCC tumor immune microenvironment, elucidate the interactions between immune cells and tumors, and establish prognostic risk model. Methods: 20 HNSCC samples single-cell rna sequencing (scRNA-seq) data and bulk rna-seq data were derived from public databases. The "cellchat" R package was used to identify cell-to-cell communication networks and prognostic related genes, and then cell-cell communication (ccc) molecular subtypes were constructed by unsupervised clustering. Kaplan-Meier(K-M) survival analysis, clinical characteristics analysis, immune microenvironment analysis, immune cell infiltration analysis and CD8+T cell differentiation correlation analysis were performed. Finally, the ccc gene signature including APP, ALCAM, IL6, IL10 and CD6 was constructed based on univariate Cox analysis and multivariate Cox regression. Kaplan-Meier analysis and time-dependent receiver operating characteristic (ROC) analysis were used to evaluate the model in the train group and the validation group, respectively. Results: With CD8+T cells from naive to exhaustion state, significantly decreased expression of protective factor (CD6 gene) is associated with poorer prognosis in patients with HNSCC. The role of macrophages in the tumor microenvironment has been identified as tumor-associated macrophage (TAM), which can promote tumor proliferation and help tumor cells provide more nutrients and channels to facilitate tumor cell invasion and metastasis. In addition, based on the strength of all ccc in the tumor microenvironment, we identified five prognostic ccc gene signatures (cccgs), which were identified as independent prognostic factors by univariate and multivariate analysis. The predictive power of cccgs was well demonstrated in different clinical groups in train and test cohorts. Conclusion: Our study highlights the propensity for crosstalk between tumors and other cells and developed a novel signature on the basis of a strong association gene for cell communication that has a powerful ability to predict prognosis and immunotherapy response in patients with HNSCC. This may provide some guidance for developing diagnostic biomarkers for risk stratification and therapeutic targets for new therapeutic strategies.

[In vivo measurement of three-dimensional motion of the upper cervical spine using CT three-dimensional reconstruction].

OBJECTIVE: Using the CT three-dimensional reconstruction to measure the activity degree of atlanto-occipital joint and the atlantoaxial joint in different directions and its coupling movement in healthy volunteers, and three dimensional motion range of the maximum rotation position of the upper cervical spine of cervical spondylosis patients, and to analyze the differences, verifing the reliability of the method at the meantime. METHODS: From January 2014 to June 2015, 20 healthy adult subjects(healthy adult group), and 26 patients with cervieal spondylosis(cervical spondylosis group) were selected. In healthy adult group, there were 11 males and 9 females, aged from 22 to 26 years old with an average of (24.0±1.2) years, and in cervical spondylosis group, there were 24 males and 2 females, aged from 36 to 72 years old with an average of (52.8±8.6) years. Healthy adults underwent CT examination in neutral position, maximum right rotation, maximum right lateral bending, maximum flexion and extention, and cervical spondylosis patients underwent CT examination in neutral position, maximum right rotation. Then the software Mimics was used to reconstruct occiput (Oc), atlas(C1) and axial(C2) vertebral three-dimensional image. Three virtual non-collinear markers were positioned on prominent structures of foramen magnum, C1 and C2. The 3D spatial coordinates of these virtual anatomical markers entail the definition of an anatomical local coordinate system which represent the position and orientation of the bones. Segmental motions were calculated using Eulerian angle in three major planes, and the difference between cervical spondylosis group and healthy adult group were compared. Due to the inaccuracy in anatomical landmark idenrification, two groups were measured 3 times, and the reliability of the experimental metnod was verified by the intra-group correlation (intra-group ICC) and the inter-group correlation coefficient(inter-group ICC). RESULTS: Reliability verification results:the intra-group ICC and inter-group ICC results were all above 0.90, and the measurement method had high reliability. Three-dimensional activity of the upper cervical spine in healthy adults:the atlanto-occipital joint had(-6.8±1.5)° coupled left lateral bending and (8.9±2.0)° coupled extension in the maximum right rotation position, and the motion of atlanto-occipital joint had low activity[maximum was(5.3±2.6)°] in the remaining 3 positions; the rotation of atlanto-axial joint was(37.9±5.1)°, accounting for 52.34% of the total cervical spine activity[(72.4±5.0)°] in the maximum right rotation position, and rotational motion was still prominent in the remaining three positions. The relative translations of the upper cervical spine in all direction were small. The average axial rotation angle [(62.0±3.4)] ° of the total cervical spine in cervical spondylosis group was significantly lower than that in the healthy adult group, but the mean axial rotation angles of the atlanto-occipital and the atlantoaxial joint were not significantly different from those of the healthy adults(P>0.05). CONCLUSIONS: The three-dimensional CT reconstruction method has high reliability, which can be applied to measure the movement of spine. The upper cervical spine contributed the most to the direction of rotation, and the movement in all directions are accompanied by coupled motion in the other direction. There was no significant difference in the rotation of the upper cervical spine between cervical spondylosis patients and normal subjects.

[Validity study of Coda Motion Analysis System for measuring cervical lateral flexion in normal adults].

OBJECTIVE: To investigate the validity of Coda Motion Analysis System for measuring cervical lateral flexion in normal adults in order to explore a new measuring tool for clinical and research practice. METHODS: A total of 43 participants were involved in the study. Cervical range of lateral flexion were measured with Coda Motion Analysis System and "gold standard" X-ray simultaneously. The validity and agreement were assessed using the scatter diagram, the Pearson correlation coefficient and limits of agreement. RESULTS: Cervical range of lateral flexion measured by Coda Motion Analysis System had no statistical differences with those measured by X-ray(P>0.05). The Coda Motion Analysis System demonstrated a very good linear relation with the X-ray measurements in cervical range of right lateral flexion, left flexion and total lateral flexion, and the Pearson correlation coefficients were 0.72, 0.85 and 0.90 respectively. CONCLUSIONS: Coda Motion Analysis System showed good validity for measuring cervical lateral flexion in normal adults. Because the reliability of Coda Motion Analysis System was established previously, the results of this study suggest that the system has the potential to be used to measure cervical lateral flexion in clinical and research practice.

Characteristics and Management of Free Flap Compromise Following Internal Jugular Venous Thrombosis.

PURPOSE: Free flap compromise after internal jugular venous thrombosis (IJVT) occurs fairly infrequently in oral and maxillofacial rehabilitation and reconstruction but could seriously affect the patient's postoperative recovery. The aim of this study was to analyze and discuss the characteristics and management of such flap compromise. PATIENTS AND METHODS: The authors performed a retrospective case series of 13 patients who underwent reconstruction of oral and maxillofacial defects with anterolateral thigh flaps and developed flap compromise from IJVT from July 2014 through December 2016 at the Second Xiangya Hospital of Central South University (Changsha, China). The clinical features of flap compromise are described, and the precautions and improvement measures are reported. RESULTS: All 13 patients were men with an average age of 50.3 years. Flap compromise caused by IJVT occurred on postoperative days 3 to 7, of which 2 flaps were salvaged (15.4%) and 11 flaps failed completely after immediate urgent re-exploration. Four patients presented with orocutaneous fistula after salvage surgery that healed well after a period of wound dressing. CONCLUSIONS: Because of the late occurrence and low salvage rate of flap compromise caused by IJVT, intensive flap monitoring after reconstruction surgery is necessary for patients with free flap transfer.

[New progress on three-dimensional movement measurement analysis of human spine].

Spinal biomechanics, especially the range of spine motion,has close connection with spinal surgery. The change of the range of motion (ROM) is an important indicator of diseases and injuries of spine, and the essential evaluating standards of effect of surgeries and therapies to spine. The analysis of ROM can be dated to the time of the invention of X-ray and even that before it. With the development of science and technology as well as the optimization of various types of calculation methods, diverse measuring methods have emerged, from imaging methods to non-imaging methods, from two-dimensional to three-dimensional, from measuring directly on the X-ray films to calculating automatically by computer. Analysis of ROM has made great progress, but there are some older methods cannot meet the needs of the times and disappear, some classical methods such as X-ray still have vitality. Combining different methods, three dimensions and more vivo spine research are the trend of analysis of ROM. And more and more researchers began to focus on vivo spine research. In this paper, the advantages and disadvantages of the methods utilized recently are presented through viewing recent literatures, providing reference and help for the movement analysis of spine.

[Effects of early supplemental parenteral nutrition on nutrition intakes and clinical outcomes in trauma patients].

OBJECTIVE: To evaluate energy and protein intake changes in early supplemental parenteral nutrition (PN) in trauma patients, and to assess its impact on clinical outcomes. METHODS: Clinical results of patients receiving or not receiving additional PN during the first 7 days after injury were retrospectively analyzed, with a total of 195 patients classified into two groups: control group (n=105) and mixed nutrition group (n=90). The time of nutrition support, intakes of protein and energy within 14 days after trauma, and clinical outcomes were compared between two groups. RESULTS: The degree of injury was comparable between two groups with no significant differences in acute physiology and chronic health evaluation II score, injury severity score (ISS) and Glasgow coma score (GCS). Compared with the control group, the mixed nutrition group received parenteral nutritional support earlier (40.0±21.0 hours vs. 55.1±23.5 hours, P<0.01), with later beginning of enteral nutrition (EN, 75.2±54.5 hours vs. 55.1±23.5 hours, P<0.01) and lower rate of EN in 48 hours after admission [14.4% (13/90) vs. 43.8% (46/105), P<0.01]. The time of restoring oral diet was not different between the mixed nutrition group and control group (10.8±3.7 days vs. 11.4±3.6 days, P>0.05). The energy intake was significantly higher in the mixed nutrition group than in the control group in 3, 7, 14 days (3 days: 3981.6±2209.3 kJ vs. 2683.2±1414.9 kJ, 7 days: 5477.5±2008.4 kJ vs. 3619.1±1429.9 kJ, 14 days: 6250.2±2533.2 kJ vs. 5199.9±1972.7 kJ, P<0.05 or P<0.01). In both groups the protein intake was insufficient, and it was significantly lower in the mixed nutrition group than in the control group on day 3 (20.6±18.4 g vs. 26.5±13.8 g, P<0.05). The patients in the mixed nutrition group had longer hospital stay time (73.9±62.5 days vs. 50.9±33.3 days, P<0.01). The mortality rate of mixed nutrition group and control group was 4.4% (4/90) and 3.8% (4/105) respectively, the rate of infection and acute respiratory distress syndrome (ARDS) were 8.9% (8/90) and 3.8% (4/105), 5.6% (5/90) and 7.6% (8/105) respectively, duration of mechanical ventilation (days) was 8.3±4.6 and 7.3±4.7, duration of stay in ICU was 17.6±13.2 days and 14.2±11.3 days respectively, and no significant difference was found between two groups (all P>0.05). CONCLUSION: Although early supplemental PN within 7 days after injury increases energy intake, PN without a standard protocol does not improve clinical outcomes and may prolong hospital stay time.

[Analysis of energy balance and risk factors on clinical outcomes in patients with severe traumatic brain injury].

OBJECTIVE: To observe the energy expenditure in severe traumatic brain injury patients, and to assess the impact of cumulative energy balance on clinical outcomes. METHODS: Using prospective self-controlled study, the change in energy expenditure of 42 patients with severe traumatic brain injury was measured by indirect calorimetry (IC). Daily energy intake was recorded. Afterwards, energy balance was calculated. The levels of nutritional biochemical indicators were compared. Logistic regression analysis was used to analyze the correlation of cumulative energy balance with clinical outcomes. RESULTS: Mean practical energy intake of all patients was (6787 ± 1848) kJ/d, and mean negative energy balance was (913 ± 285) kJ/d. The negative energy balance was most crucial in first 3 days after admission. Meanwhile, practical energy intake was significantly lower than target energy intake (kJ: 2859 ± 1370 vs. 6027 ± 899, P < 0.01). The practical energy intake was increased with time, and it was found that the first 14 days were crucial for development of negative energy balance. On 7th day after admission, albumin (g/L) level in plasma was lowest compared with that on 3rd day (29.5 ± 5.0 vs. 35.9 ± 3.8, P < 0.01), and then it was increased gradually returning to normal level on 28 days (34.1 ± 2.8). Three days after admission, prealbumin (mg/L: 122.5 ± 23.3) was obviously lower than normal level, but it rapidly elevated on 7th day (214.3 ± 38.6, P < 0.01) and continued to rise till 28th day (257.7 ± 25.2). On the 3rd day after admission, C-reactive protein (mg/L: 139.5 ± 54.4) was obviously higher than normal level. However, it significantly fell on 7th day (108.4 ± 42.2, P < 0.01), and it continued to fall. Logistic regression analysis showed a strong association of cumulative negative energy balance with infection and upper gastrointestinal bleeding [odds ratio (OR) of infection was 2.130, 95% confidence interval (95%CI) 1.540 to 29.661, P = 0.023; OR of upper gastrointestinal bleeding was 0.083, 95%CI 0.013 to 0.542, P = 0.009]. CONCLUSIONS: Cumulative negative energy balance may be correlated with the occurrence of complications in patients with severe traumatic brain injury. On the basis of the measurements of changes in energy by IC, early supply of sufficient energy may improve the outcome of patients.