Objective:To investigate the clinical application value of plasma SEPT9 gene methylation combined with serum carcinoembryonic antigen (CEA) and carbohydrate antigen 724 (CA724) in the diagnosis of colorectal cancer.Methods:A total of 219 patients with colorectal diseases in Baoji Central Hospital and Yunnan Province New Kun Hua Hospital from May 2018 to October 2021 were selected, including 149 cases of colorectal cancer and 70 cases of colorectal polyp diagnosed by pathology. A total of 100 healthy people in the same period were selected as the healthy control group. The methylation of SEPT9 gene in plasma was measured by using real-time fluorescent polymerase chain reaction (PCR), and the levels of serum CEA and CA724 were measured by using electrochemiluminescence. The expressions of three indicators in each group were compared, and the effect of every single indicator and the combination of the three indicators on the diagnosis of colorectal cancer was analyzed by using receiver operating characteristic (ROC) curve.Results:The positive rate of SEPT9 gene methylation in colorectal cancer group (74.50%, 111/149) was higher than that in colorectal polyp group (22.86%, 16/70) and healthy control group (1.00%, 1/100), and the difference was statistically significant ( P < 0.001). The positive rate of CEA in colorectal cancer group (46.98%, 70/149) was higher than that in colorectal polyp group (40.00%, 28/70) and the healthy control group (3.00%, 3/100) and the difference was statistically significant ( P < 0.001). The positive rate of CA724 in colorectal cancer group (38.93%, 58/149) was higher than that in colorectal polyp group (32.86%, 23/70) and the healthy control group (2.00%, 2/100), and the difference was statistically significant ( P < 0.001). The area under the curve (AUC) of ROC of SEPT9 gene methylation, CEA and CA724 in the single diagnosis of colorectal cancer was 0.823 (95% CI 0.753-0.891), 0.788 (95% CI 0.725-0.852) and 0.689 (95% CI 0.624-0.754), respectively. The optimal cut-off Ct value of SEPT9 gene methylation in the diagnosis of colorectal cancer was 36.5, the sensitivity was 90.30%, and the positive predictive value was 84.68%, which were higher than those of CEA and CA724. The optimal cut-off value of CEA in the diagnosis of colorectal cancer was 8.80 ng/ml, and the specificity (77.50%) and negative predictive value (78.48%) were higher than those of SEPT9 gene methylation and CA724. The sensitivity (97.66%), positive predictive value (93.98%), negative predictive value (81.25%) and AUC (0.846, 95% CI 0.749-0.944) of the combined detection of the three indexes taking the optimal cut-off value of every single indicator were higher than those of the single indicator. Conclusions:The combined detection of plasma SEPT9 gene methylation, CEA and CA724 in the diagnosis of colorectal cancer has high sensitivity and accuracy. The three combined detection can complement each other and improve the diagnostic efficiency, which is of high clinical value for the diagnosis of colorectal cancer.

Objective: To investigate the mechanism of lncRNA HCG18/miR-17-5p/HMGA2 axis regulating the proliferation and metastasis of non-small cell lung cancer (NSCLC) cells. Methods: Sixty-two pairs of NSCLC tissues and corresponding para-cancerous tissues collected at Central Hospital of Chengde City from June 2017 to June 2018 were used for this study; in addition, NSCLC cell lines (A549, NCI-H1299, H1650, NCI-H460) and human lung epithelial BEAS-B cells were also collected. mRNA expression levels of HCG18, miR-17-5p and high-mobility group AT-hook 2 (HMGA2) in NSCLC tissues and cell lines were measured by quantitative Real-time polymerase chain reaction (qPCR). Si-HCG18, miR-17-5p, miR-17-5p+HCG18 or pcDNA3.1-HMGA2 were transfected into A549 cells and NCI-H460 cells; CCK-8 assay was used to detect the proliferation of transfected cells, Transwell assay was used to detect the migration and invasion ability of cells, and Wb was used to analyze the expressions of HMGA2 and EMT associated proteins (E-cadherin, N-cadherin and vimentin). The target relationships between HCG18 and miR-17-5p, or between miR-17-5p and HMGA2 were confirmed by dual luciferase reporter gene assay. Mice A549 cell xenograft model with HCG18 knockdown was constructed, and the growth of transplanted tumor was observed. Results: lncRNA HCG18 was highly expressed in NSCLC tissues and cells (all P<0.01); HCG18 level was significantly increased in patients at late stage or with lymphnode metastasis; and high HCG18 level was correlated with poor prognosis and low survival rates of NSCLC patients (all P<0.01). Knockdown of HCG18 significantly inhibited NSCLC cell proliferation, migration and invasion (all P<0.01), up-regulated E-cadherin expression but suppressed N-cadherin and vimentin expression (all P<0.01), and the volume of xenograft was obviously decreased (P<0.05). Dual luciferase reporter gene assay confirmed the relationship between HCG18 and miR-17-5p as well as miR-17-5p and HMGA2. miR-17-5p transfection significantly inhibited NSCLC cell proliferation, migration and invasion (all P<0.01), and up-regulated E-cadherin expression, reversely suppressed N-cadherin and vimentin expression (all P<0.01); however, miR-17-5p + HCG18 transfection reversed the effect of miR-17-5p on NSCLCcells.Conclusion:HCG18promotes the proliferationandmigrationofNSCLCcellsthrough regulating miR-17-5p/HMGA2 axis.

OBJECTIVETo explore the detection efficiency of circulating tumor cells (CTCs) in patients with hepatocellular carcinoma (HCC).

METHODSImmunomagnetic negative enrichment by nanometer magnetic beads and label-free capture with Captor(TM) system were used to isolate and enrich CTCs from peripheral blood of HCC patients, and epithelial and HCC markers were applied to identify CTCs by immunofluorescence staining. CTCs were detected in 50 HCC patients before and after hepatectomy to test the method for isolation, enrichment and identification. The dynamic changes of pre- and post-operative CTCs' numbers were compared. The clinical data were analyzed using SPSS 19.0 software.

RESULTSNegative enrichment methods by nanometer magnetic beads and label-free capture using Captor(TM) system were both suitable for CTCs isolation and enrichment in HCC patients. The positive detection rate of CTCs in HCC patients via negative enrichment was 96.0% (48/50), the preoperative median number of CTCs was 16 per 7.5 ml blood, and the postoperative median number was 17 per 7.5 ml blood.

CONCLUSIONSBoth negative enrichment and Captor(TM) system are suitable for isolation and enrichment of CTCs in HCC patients. There is a significant difference in the numbers of CTCs before and after operation, and dynamic detection of CTCs will provide helpful prognostic information for HCC patients in clinics.

Adult ; Aged ; Antigens, Neoplasm ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Adhesion Molecules ; metabolism ; Epithelial Cell Adhesion Molecule ; Female ; Hepatectomy ; Humans ; Immunomagnetic Separation ; methods ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; metabolism ; pathology

Objective To explore the characteristic of sleep disorders in the initial stage of withdrawal and their relationships with alcohol craving levels in alcohol dependence (AD) patients,and provide support for diagnosis and prevention of re-drinking.Methods Thirty-two AD inpatients were assigned to AD group and 20 male healthy volunteers to control group.Alcohol craving was assessed with the Pennsylvania Alcohol Craving Scale (PACS) within the 2nd week after alcohol withdrawal for AD patients,and then the whole-night polysomnogram (PSG) tracings were conducted on the day of the night.Results The five item scores of the PACS were from 3.48 to 4.26 in AD patients.The sleep latency was(42.48±22.42) min,total sleep time was(289.61± 103.22)min,sleep efficiency was(71.45± 19.86) ％,S1 sleep was (23.47± 11.07) ％,arousal frequencies was (8.01 ± 2.77),S3+4 sleep was(6.26±5.35)％ in AD group.Compared with control group((19.65±8.57) min,(407.33±21.29) min,(81.52 ± 6.46) ％,(8.79± 1.83) ％,(2.17 ± 1.04),(15.87 ± 5.24) ％ respectively),the differences had statistical significances(t=2.206-9.082,P＜ 0.05-0.001).Alcohol craving levels were positively related to sleep latency,arousal frequencies and S1 sleep (r=0.424-0.898,P＜0.05-0.01) and negatively to total sleep time,sleep efficiency and S3+4 sleep (r=-0.416--0.662,P＜0.05-0.01) in AD group.Conclusion AD patients have sleep continuity and structure disturbances in the initial stage of alcohol withdrawal,sleep continuity and structure disturbances are related to alcohol craving.Improvements of sleep disorders should be paid during clinical alcohol dependence treatment.

Objective To study the reasons and prevention of the hoarse voice after continuity-intact recurrent laryngeal nerve (RLN)operation without damage of RLN under naked eyes.Methods Data of 1871 patients undergoing thyroid surgery from Jan.2001 to Jan.2011 were retrospectly analyzed.919 patients had their RLN exposed,among whom 757 patients had bilateral RLN exposed.952 patients didn't have their RLN exposed in the surgery.Results A total of 1676 RLNs were exposed by the routine method and minimally invasive method.All the nerves were confirmed no damage and continuity intact under naked eyes.Hoarse voice occurred to 19patients after surgery,with the percentage of 2.12％ (19/897).The rate of hoarse voice in the non-exposed group was 5.46％ (52/952).The difference of horse voice between the RLN exposed group and non-exposed group had statistical significance(P ＜ 0.05).Conclusions RLN without damage under naked eyes and continuity intact doesn't mean no damage of electroneurophysiology.The rate of RLN damage in the exposed group was less than that in the non-exposed group.The major causes of hoarse voice may include misoperation,heat conduction and postoperative scar adhesion.The key to avoid RLN damage is prevention.

Objective To explore the effect of endocrine therapy in breast cancer patients whose estrogen receptor (ER) and progesterone receptor(PR) was preoperatively negative and turned positive after neoadjuvant chemotherapy. Methods The clinical experimental study was carried out in 97 cases of breast cancer which were divided into endocrine treatment group and control group. The follow-up time ranged from 15 to 60 months. Results In endocrine treatment group, 3 and 5-year disease-free survival were respectively 74.5% (38/51), 60.7% (31/51), and 3 and 5-year overall survival were respectively 80%(41/51), 74. 5% (38/51). In control group, 3 and 5-year disease-free survival were respectively 54.2% (26/46), 41.7%(20/46), and 3 and 5-year overall survival were 60.9%(28/46),50%(23/46),respectively. The corresponding values were significantly higher in endocrine treatment group than in control group(P＜0.05).Conclusions Remedy endocrine therapy improves the disease-free and overall survival rate in breast cancer patients with the expression of ER and PR turning positive after initial neoadjuvant chemotherapy.