Objective To investigate the role of NMDA receptors in central medial thalamus (CMT) in the unconscious?ness induced by general anesthesia. Methods A total of 60 rat models for microinfusion were assigned into 4 groups (n=15 for each group). After induction with propofol, 10 mmol/L (NMDA10 group), 20 mmol/L (NMDA 20 group) and 40 mmol/L (NMDA40 group) of NMDA and normal saline (group C) with equal volume were microinfused into CMT. The incidence of purposeful movement and recovery time of righting reflex were observed in each group respectively. Infusion sites were local?ized by histological method. Results When the microinfusion site localized within CMT, comparing with group C, the recov?ery time of righting reflex reduced notably in three NMDA groups (P<0.05). The recovery time was significantly shorter in NMDA20 group and NMDA40 group than that of NMDA10 group. The incidence of purposeful movement during propofol an?esthesia was higher in NMDA20 group and NMDA40 group than that of group C (P<0.05). When the microinfusion site lo?calized out of CMT, the recovery time of righting reflex was remarkably longer than that within CMT in three NMDA groups (P<0.05), and there was no significant difference in the incidence of purposeful movement and recovery time between four group (P>0.05). Conclusion Microinfusion of NMDA agonist into CMT reverses propofol anesthesia, indicating that NMDA receptor in CMT may contribute to the propofol-induced unconsciousness.

OBJECTIVETo detect the expression levels of programmed death-1 (PD-1) and programmed death ligand-1 (PD-L1) in the peripheral blood of patients with oral squamous cell carcinoma (OSCC) and to discuss their biological and clinical significance.

METHODSPD-1/PD-L1 expression on the surface of T-lymphocytes and the counts of T-lymphocyte subpopulations of peripheral blood in 82 patients with OSCC (OSCC group) and 25 healthy controls (control group) were examined via flow cytometry. The expression levels of soluble PD-1 (sPD-1) and soluble PD-L1 (sPD-L1) in the serum were observed through enzyme-link immunology method. The data were tested and analyzed with SPSS 17.0 software.

RESULTSThe percentage of CD8+ T cells in the OSCC group was significantly higher than that in the control group (P<0.05), whereas the percentages of CD3+ and CD4+ T cells as well as CD4+/CD8+ ratio were significantly lower than those in the control group (P<0.05). The positive rates of PD-1 and PD-L1 in CD3+ and CD4+ T cells in OSCC peripheral blood were remarkably higher than those in the control group (P<0.01). Difference was not observed between the expression levels of sPD-1 in the serum of OSCC group and those in the control group (P>0.05), but the average of sPD-L1 was remarkably higher than that in the control group (P<0.05). sPD-L1 expression was related to clinical stage, tumor cell differentiation, and lymph node status (P<0.05) but not related to sex, age, tumor location, and tumor size.

CONCLUSIONT-lymphocyte subpopulations in the peripheral blood of patients with OSCC developed immunosuppression with different degrees. PD-1 and PD-L1 expression levels on the surface of CD3+ and CD4+ T cells significantly increased. Abnormal increase in sPD-L1 expression may be associated with OSCC development.

B7-H1 Antigen ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; Case-Control Studies ; Flow Cytometry ; Humans ; Mouth Neoplasms ; metabolism ; T-Lymphocyte Subsets

Objective To construct a recombinant bacterial vaccine which can express specific 10-23 deoxyribozyme(DZ) in macrophage, identify the intracellular production of specific 10-23DZ and detect the activity of this recombinant bacterial vaccine on inhibiting the expression of TACO gene in macrophage.Methods The pSDE02 was obtained by inserting the replicon of Mycobacterium into pSDE01, a plasmid which can express 10-23DZ in eukaryotic cells. The expression sequence of DZ1, a 10-23DZ targeting the TACO mRNA of macrophage designed in our previous study was synthesized and inserted into pSDE02. The resulted plasmid was named pDZM01. pDZM01 was then transferred into Mycobacterium smegmatis by electroperation. The recombinant M. smegmatis, named rMs-DZ1 was screened on low-salt LB medium containing Zeocin and identified by Colony PCR. The targeted delivery property of recombinant M. smegmatis was observed by Ziehl-Heelson stain and GFP expression observation via fluorescence microscope. rMs-DZ1 was used to infect RAW264.7 cells and the expression of DZ1 in macrophage was identified by dot-blot assay. At 24 h and 48 h after infection, total RNA and proteins were extracted and the TACO mRNA and protein expression level was assayed by RT-PCR and western-blot respectively. Results Restrictive analysis and sequencing data showed that the Mycobacterium-eukaryotic cell shuttle plasmid pSDE02 and pDZM01 was successfully constructed. rMs-DZ1 was confirmed by colony PCR. When engulfed by macrophage, rMs-DZ1 would express DZ1 in RAW264.7 cells and inhibit the expression of taco gene. When compared to uninfected macrophage, rMs-DZ1 significantly reduced the taco mRNA by 67.90% and 57.14% and down-regulated the expression of TACO protein by 53.85% and 68.92% at 24 h and 48 h respectively. Conclusion A recombinant M. smegmatis vaccine was successfully constructed which could generate specific 10-23DZ in macrophage and inhibit the expression of target gene of interest. To our knowledge, this is the first bacterial vector which can express intracellularly 10-23DRz in targeted manner. This study may further prompt the feasibility of using 10-23 DNAzyme to achieve effective and targeted gene silence.

Objective To retrospectively analysis and compareabout Bryan artificial cervical disc arthroplasty with ante-rior cervical decompression and fusion (ACDF) on the clinical efficacy for“Skip”cervical spondylosis. Methods From February 2002 to May 2012, 49 cases were treated with Bryan artificial cervical disc arthroplasty (artificial cervical disc replacement surgery group, 18 cases) or anterior cervical decompression and fusion (ACDF group, 31 cases), 29 males and 20 females. Each case was evaluated at the moment of preoperatively, 3 months, 6 and 12 months and last follow-up after surgery by the Japanese Orthopedic Association (JOA), Neck Disability Index (NDI), Visual Analog Scale (VAS), Cervical sagittal curvature, the total cervical spine range of motion(ROM),middle segments of motion. MRI was also used to assess to adjacent segment disc degeneration, spinal cord compression and signal change situation. Results All patients were followed up for more than 24 months. The score of the JOA, NDI, VAS in the two groups of patients improved significantly after surgery than before surgery. In addition, the VAS score in last follow-up were significantly different between the two groups, but other index each time in the two groups showed no significant difference. In last follow-up, the result of artificial cervical disc arthroplasty group were better than ACDF group on the incidence of axial symptoms, the total cervical spine range of motion (ROM) and middle segments of motion. The incidence of axial symptoms in artificial cervical disc arthroplasty group were 11.1%,ACDF group were 45.2%. ROM in arti-ficial cervical disc arthroplasty group were 35.5°±5.9°,ACDF group were 24.5°±6.2°. Middle segments of motion in artificial cer-vical disc arthroplasty group were 7.3°±1.4°,ACDF group were 10.1°±1.6°. The above comparison of the datas were statistically different. There are two cases of adjacent segment degeneration in ACDF group without need to surgery. Conclusion Bryan artifi-cial cervical disc replacement surgery effectively retained the overall motion of the cervical spine, reduced the motion of middle segments, thus avoiding adjacent segment degeneration and the incidence of postoperative axial symptoms.

Background:With the improvement of national health awareness and the popularization of a series of screening methods,the number of early colorectal cancer is gradually increasing,accurate prediction of lymph node metastasis of T1 colorectal cancer is the key to determine the optimal therapeutic solutions.Aims:To investigate the relationship between preoperative inflammatory markers,fibrinogen,apolipoprotein B and lymph node metastasis in patients with early colorectal cancer.Methods:The clinical data of 102 patients with early colorectal cancer who received surgical treatment in the Department of General Surgery,Affiliated Hospital of Jiangsu University from January 2014 to December 2022 were retrospectively analyzed.The patients were divided into positive lymph node group and negative lymph node group according to postoperative pathological results.Univariate and multivariate Logistic regression analysis were employed to explore the correlation between lymph node metastasis and clinical test parameters in early colorectal cancer.Results:A total of 102 patients in T1 colorectal cancer were enrolled in this study,including 53 males and 49 females,and the mean age was(64±10)years.Postoperative pathological diagnosis of lymph node metastasis was 13 cases and no lymph node metastasis was 89 cases.The univariate analysis showed that age,fibrinogen and apolipoprotein B-monocyte ratio(AMR)were related to lymph node metastasis in early colorectal cancer(P<0.05).The multivariate Logistic regression analysis showed that age,fibrinogen and AMR were independent predictors of lymph node metastasis.Conclusions:The age,fibrinogen and AMR may play an important role in predicting lymph node metastasis in early colorectal cancer.They can be combined with pathological factors to further create a new prediction model,so as to provide some reference for patients in colorectal cancer to choose the therapeutic regimen.

OBJECTIVETo investigate the expression of long non-coding RNA(lncRNA) colon cancer associated transcript 2(CCAT2) and its association with clinicopathologic features in oral squamous cell carcinoma(OSCC).

METHODSThe expression of lncRNA was detected with microarray assay in three samples of OSCC tumor and matched adjacent tissues. The profiles of lncRNAs in OSCC tissues were identified. The CCAT2 expression was evaluated by real-time quantitative PCR(RT-qPCR) in 86 OSCC tumor samples and matched adjacent tissues. The relationship between the expression of CCAT2 and its clinicopathologic features of OSCC was analyzed. Tumor cell proliferation was assessed following siRNA knockdown of CCAT2 by using the CCK-8 kits.

RESULTSA total of 1 685 lncRNA expressed in OSCC tumor samples and matched adjacent tissues were identified using microarray assay(P<0.05). RT-qPCR showed that the expression of CCAT2 was significantly higher in OSCC than that in adjacent tissues(P< 0.01). High CCAT2 expression was associated with cell differentiation and pathological stage of OSCC. CCAT2 expression in low-differentiated OSCC was significantly higher than that in high-differentiated cancer (P=0.015). In addition, CCAT2 level in stage Ⅲ/Ⅳ OSCC was significantly higher than that in stage Ⅰ/Ⅱ cancer (P=0.022). Furthermore, inhibition of CCAT2 expression suppressed the proliferation of human tongue carcinoma Tca8113 cells.

CONCLUSIONSAbnormal expression of lncRNA may be involved in the development of OSCC. Up-regulation of CCAT2 expression in tumor tissue might act as an oncogene and promote the development of OSCC.

Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Proliferation ; Gene Expression Profiling ; Humans ; Mouth ; metabolism ; Mouth Neoplasms ; genetics ; pathology ; RNA, Long Noncoding ; analysis ; metabolism ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Up-Regulation

Objective : To analyze circular RNA (circRNA) expression profiles in oral squamous cell carcinoma (OSCC) and its clinical significance. Methods : The expression of circRNA was detected with circRNA microarray assay in three samples of OSCC tumor and matched adjacent tissues. Quantitative real-time PCR (RT-qPCR) was used to verify the expression of circRNA in 45 pair OSCC tissues and normal adjacent tissues. The relationship between the expression of circRNA and the clinicopathological characteristics of OSCC was analyzed. circRNAs/miRNAs interaction were predicted using Arraystar' s home-made miRNA target prediction software. Results : 155 circRNAs were differentially expressed between the OSCC tissues and matched adjacent tissues, of which 45 circRNAs were up-regulated and 110 circRNAs were down-regulated in OSCC tissues (fold changes ≥1.5 and P < 0.05). In the selected three circRNAs that were most significantly upregulated or downregulated in OSCC, the RT-qPCR results showed that hsa_circ_0001874, hsa_circ_0001971 and has_circ_0067934 were increased, while hsa_circ_0000520, hsa_circ_0023944 and hsa_circ_0000140 were decreased in OSCC tissues versus normal adjacent tissues (P < 0.001). The results were generally consistent with the microarray data. Among the circRNA expression profiles in OSCC, the up-regulation of hsa_circ_0001874 was the highest and the expression of hsa_circ_0001874 was significantly correlated with TNM stage and tumor grade. The result of Arraystar' s home-made miRNA target prediction software indicated that miR-103a-3p, miR-107, miR-593-5p, miR-661 and miR-662 may be potential target genes of hsa_circ_0001874. Conclusion The differentially expressed circRNAs in OSCC tissues and normal adjacent tissues were identified, and these dysregulated circRNAs and their potential target gents may play important roles in the development of OSCC.

Objective: To explore the expression and clinical significance of circular RNA circHIPK3 in oral squamous cell carcinoma (OSCC), analyze the effect of circHIPK3 on the proliferation of OSCC cells. Methods: The expression of circHIPK3 in OSCC tissues, adjacent non-cancerous tissues and OSCC cell lines were detected by quantitative real-time polymerase chain reaction (qPCR). The correlations between the expression of circHIPK3 in OSCC tissues and the clinicopathological features were analyzed as well. circHIPK3-specific siRNA si-circHIPK3 and negative control siRNA si-NC were designed and synthesised and used to transfect CAL27 and SCC15 cells respectively. The proliferation capacity of CAL27 and SCC15 cells after transfection with si-circHIPK3 was detected by CCK-8 assay. The expression of miR-124 in OSCC was detected by qPCR, and the correlation between expression of circHIPK3 and the expression of miR-124 was analyzed. Using qPCR to detect the expression of miR-124 in CAL27 and SCC15 cells after transfection with si-circHIPK3 and si-NC respectively. Furthermore, using CCK-8 assay to detect the proliferation capacity of CAL27 and SCC15 cells after transfection with si-NC, si-circHIPK3, miR-124 mimic, si-circHIPK3+miR-124 inhibitor. Results: The expression of circHIPK3 in OSCC tissues [2.23 (1.86, 3.00)] was significantly higher than that of the adjacent non-cancerous tissues [1.05 (0.85, 1.26)] (U=1 094, P=0.000). The expression of circHIPK3 in CAL27 (3.02±0.51) and SCC15 cells (3.16±0.75) was higher than those of human normal oral keratinocytes (hNOK) (1.26±0.30) (P=0.000). The expression of circHIPK3 was found to be closely associated with TNM stage (P<0.05) and tumor grades (P<0.05). Knockdown of circHIPK3 can inhibit proliferation of CAL27 and SCC15 cells (P<0.05). The expression of miR-124 in OSCC tissues (0.61±0.35) was significantly lower than that in adjacent non-cancerous tissues (1.13+0.39) (t=-5.36, P<0.05). Correlation analysis showed that the expression of circHIPK3 in OSCC was negatively correlated with the expression of miR-124 (r=-0.767, P<0.001). Moreover, down-regulation of miR-124 rescued the phenotype induced by knockdown of circHIPK3. Conclusions The expression of circHIPK3 in OSCC was increased, and silencing of circHIPK3 expression can inhibit the proliferation of OSCC cells. Our results suggest that circHIPK3 may play a key role in the occurrence and development process of OSCC through the regulation of miR-124 expression.