Objective To research the effect of the Kiss-1 gene promoter methylation on the Kiss-1 gene expression in colorectal carcinoma tissue,and to analyze the relationship between the Kiss-1 gene methylation and the clinical pathological features of colorectal carcinoma and its clinical significance.Methods The Kiss-1 gene promotor region methylation,Kiss-1 gene mRNA and protein expressions were detected respectively by methylation-specific PCR, Real-time PCR and Western blotting method in 126 cases of colorectal carcinoma tissue and para-cacinoma normal colorectal tissue.Results The positive rate of Kiss-1 gene methylation in colorectal carcinoma tissue (83.33%)was significantly higher than that in normal tissue (30.16%)(P<0.05).In the cancer tissue,the expression levels of Kiss-1 gene mRNA and protein in Kiss-1 gene promoter methylation positive group was lower than that in Kiss-1 gene promoter methylation negative group (P < 0.05 ). The positive rate of Kiss-1 gene promoter methylation in T3+T4 stage group was higher than that in T1+T2 stage group (P<0.05),but the Kiss-1 gene mRNA and protein expression levels were lower than those in T1+T2 stage group (P<0.05).In the poorly and moderately differentiated group,the positive rate of Kiss-1 gene promoter methylation was higher than that in well differentiated group (P<0.05 ), but the mRNA expression level of Kiss-1 gene was lower than that in well differentiated group (P<0.05 ). In lymph node metastasis group, the positive rate of Kiss-1 gene promoter methylation was higher than that in lymph node negative group (P<0.05),but the mRNA and protein expression levels were lower than those in lymph node negative group (P<0.05 ). And the positive rate of Kiss-1 gene promoter methylation in distant metastasis group was higher than that in non-distant metastasis group (P<0.05), but the mRNA and protein expressions levels were lower than those in non-distant metastasis group (P<0.05). There were no significant associations between the positive rate of Kiss-1 gene promoter methylation and the gender, age,tumor location, and tumor diameter of the patients (P>0.05 ). Conclusion The Kiss-1 gene promoter methylation in colorectal carcinoma tissue is associated with the Kiss-1 gene expression level and the malignant characteristics of colorectal carcinoma;Kiss-1 gene promoter methylation may be used as a reference indicator for predicting the risk of metastasis of colorectal carcinoma.

Objective To study clinical effect after laparoscopic abdominal preperitoneal inguinal hernia repair methodwithout stapler. Methods 80 cases of inguinal hernia(hospitalized from February 2015 to January 2017)were divided into two groups according to the random number table method ,with 40 patients in each group. Traditional laparoscopic peritoneal inguinal hernia repair method was applied in the control group. Free stapler group received free stapler laparoscopic preperitoneal inguinal hernia repair treatment method. Operation time , amount of bleeding during surgery , the average hospitalization time after operation , the total cost of hospitalization,postoperative pain score,postoperative recovery activities time,patients′satisfaction,operation effusion after operation occurred scrotal hematoma and other complications were comparedbetween the two groups of patients. Results In free stapler group,patients′ satisfaction rate was significantly higher than the control group (P < 0.05). Area of effusion,scrotal hematoma and other complications infree stapler group after surgery were significantly lower than the control group (P < 0.05). Total cost of hospitalization,postoperative pain score, postoperative recovery time activities in free stapler group were significantly better than those in the control group (P<0.05). Operation time,bleeding amount during the operation,average hospitalization time difference between the two groups were not statistically significant(P > 0.05). Conclusion Operation time and amount of bleeding were similar between traditional laparoscopic transabdominal preperitoneal inguinal hernia repair method and free stapler in laparoscopic transabdominal preperitoneal inguinal hernia repair.Clinical effect of free stapler in laparoscopic transabdominal preperitoneal inguinal hernia repair proves to be effective with less complications ,less pain, faster postoperative recovery, and can reduce the cost of treatment.Free stapler in laparoscopic transabdominal preperitoneal inguinal hernia repair has satisfactory cosmetic results and was well received by patients,worthy of promotion.

Objective Onto investigate the indoor quality control method for qualitatively detecting the laboratory indicators of TORCH infection (rubella virus IgG ,cytomegalovirus IgG and IgM ,toxoplasma IgG and IgM ) .Methods The statistical method , normal distribution data ,ratio and standard deviation of positive rate detected by the ELISA method were adopted ,1+2s was set as the out of control rules ,the semi Lerey‐Jennings quality control chart was drawn;the direct probability calculation method was a‐dopted for the non‐normal distribution data and small probability event .The testing data of 57 batches were retrospectively ana‐lyzed .Results The positive rate of rubella virus IgG was 86 .66% ,cytomegalovirus IgG/IgM positive rates were 98 .87% and 0 .13% ,toxoplasma gondii IgG/IgM positive rates were 2 .43% and 1 .71% ,the data of 151 ,3 ,5 ,176 ,27 samples had the critical value range of five indicators .The number of out of control was once for cytomegalovirus IgG ,once and 4 times for Toxoplasma gondii IgG/IgM .Conclusion The indoor quality control for the ELISA qualitative detection of TORCH infection can adopt the data of daily detection positive rate or negative rate for monitoring the false positive .The critical value range of specimens should be fur‐ther conducted the recheck or confirmation experiment .

Objective To characterize 15 months later the quality of life (QOL) and depression of those with spinal cord injury (SCI) suffered during the Wenchuan earthquake,and to identify the influence factors.MethodsA sample of 27 individuals who sustained SCI during the Wenchuan earthquake were surveyed using face- to-face interviews.QOL was measured using the brief version of the World Health Organization's Quality of Life Assessment instrument.Depression was measured using the patient health questionnaire depression module 9.Stepwise multiple linear regression was employed to determine the factors most closely related with QOL and depression. ResultsStepwise linear regression showed a significant correlation of overall QOL with literacy and age.The younger or illiterate victims showed a lower overall QOL.General health was also associated with ability in the activities of daily living and with pain.Victims with better daily ability and less pain may have had better general health.Depression correlated with literacy and secondary condition.Illiterate patients or patients influenced more by a secondary condition displayed more serious depression than the others.Additionally,social participation was significantly associated with the physical health,psychological health and social relationships domains of QOL.ConclusionThe QOL of long-term SCI victims caused by the earthquake is closely related to age,literacy,pain,participation and physical independence.Depression is associated with illiteracy and secondary conditions.Attention should be paid to these correlations in designing long-term rehabilitation strategies.

Objective:To explore the effects of KISS1 gene transfected by lentivirus on the proliferation,invasion and migration abilities of the colorectal cancer HCT116 cells,and to clarify their mechanisms.Methods:The human colorectal cancer cells with the lowest expression level of KISS1 gene were selected.The lentiviral vectors were builted and transfected the KISS1 gene,and the cells were divided into control group (treated with PBS),empty vector group (treated with empty vector) and over-expression group(treated with KISS1 gene vector).The multiplicity of infection (MOI) of the cells was detected by fluorescence microscope.Real-time PCR and Western blotting methods were used to detect the expression levels of KISS1 mRNA and protein(metastin);CCK-8 method was used to detect the proliferation ability of the cells;Transwell chambers method was used to detect the invasion and migration abilities of the cells.Results:Among LoVo,SW620,SW480,HCT-116,and HT29 cells,the expression levels of KISS1 mRNA and protein were lowest in HCT116 cells,so they were chosen as the research carrier.After transfected with lentiviral vectors,the HCT116 cells could stably express the enhanced green fluorescent protein(EGFP) gene,and the MOI was over 80%.Compared with control group and empty vector group,the expression levels of KISS1 mRNA and protein in the cells in over-expression group were significantly increased (P<0.05);the proliferation abilities of the cells in over-expression group were decreased (P<0.05);the invasion ability and migration ability of the cells in over-expression group were decreased (P<0.05).But the differences of proliferation ability,invasion ability and migration ability of the cells between control group and empty vector group were not statistically significant (P>0.05).Conclusion:The KISS1 gene transfected by lentivirus vector can over-express KISS1 protein and inhibit the proliferation,invasion and migration abilities of the colorectal cancer cells,and the mechanism may be related to the expression of KISS1 protein.

Objective To investigate the epidemiology of adults acute viral gastroenteritis in Shanghai Changning district. Methods All of 1 554 stool specimens of adults acute gastroenteritis in Shanghai Changning district from June 2010 to December 2013, and the enzyme-linked immunosorbent assay and multiple polymerase chain reaction was used to detecte different viruses. Results In all of 1 554 cases, the average age was (46.19 ± 15.59) years. Among them, 691 persons were male, 863 persons were female. Virus infection was detected in 407 cases, and the detection rate was 26.19%. Among them, 395 cases (97.05%) were single virus infection, and 12 cases (2.95%) were mixed infection. The peak of epidemic was from every November to next February. The incidence of watery diarrhea, vomiting and fever in virus positive group was significantly higher than that in virus negative group:95.09%(387/407) vs. 88.14%(1 011/1 147), 31.20%(127/407) vs. 18.83%(216/1 147), and 11.06%(45/407) vs. 7.59%(87/1147), P<0.01 or<0.05. Conclusions Rotavirus infection is common in adults with acute viral gastroenteritis. Patients with positive virus infection had a higher incidence of watery diarrhea, vomiting and fever. The peak of epidemic is winter.

Objective To observe the effect of STK15 gene silencing on the growth of gastric cancer cell line-MKN45 in vitro and vivo. Methods STK15 expression was inhibited by RNAi techenique, STK15 protein level was detected by Western blot, the ability of MKN45 invasion in vitro was assessed by cell migration and invasion assay, the change of cell cycle distribution was detected by flowcytometry, MKN45 proliferation was measured by MTT method, and MKN45 cells treated with STK15 siRNA were transplanted subcutanuously in nude mice and their tumorgenesis ability were observed. Results After treatment with STK15 siRNA, STK15 protein level decreased obviously. Compared with control group, STK15- group showed lower invasion ability in vitro [ mean A value: (182?27 ) vs. (308?38 ), P

Objective To explore the relationship of UHRF1 with the clinicopathological characteristics of colorectal cancer (CRC) patients, as well as the effects of lentivirus transfection overexpression and knockdown of UHRF1 on the proliferation, invasion, and migration of CRC cells and the possible signaling pathways. Methods The expression of UHRF1 mRNA and protein in CRC tissues and adjacent tissues was detected by immunohistochemical staining and RT-PCR. The effects of the constructed UHRF1 overexpression- and knockdown-group cells on the expression of UHRF1, related molecules in the WNT signaling pathway, and MMPR9 were examined by Western blot and RT-PCR. EDU and Transwell assays were used to detect changes in the proliferation, migration, and invasion of CRC cells. Results (1) In the TCGA database and clinical data, the mRNA and protein expression levels of UHRF1 in CRC cancer tissues were significantly higher than those in adjacent normal tissues. UHRF1 expression was closely correlated with TNM stage, N stage, and M stage. Patients with low UHRF1 expression in TCGA had better 5-year OS and disease-specific survival. The area under the ROC curve of UHRF1 for predicting 1-, 3-, and 5-year OS were 0.634, 0.652, and 0.771, respectively. The 3-year OS in the clinical data also showed the same survival benefit. UHRF1 overexpression was a poor prognostic factor for CRC patients. (2) After UHRF1 overexpression, the expression of WNT3a, GSK3β, and MMP9 in SW480 cells significantly increased, whereas the expression of p-β-catenin decreased (P < 0.05). After UHRF1 knockdown, the expression of WNT3a, GSK3β, and MMP9 in HCT116 cells decreased, whereas the expression of p-β-catenin increased (P < 0.05). The "rescue" experiment with IWP-2 and HLY78 can produce consistent results. (3) Compared with the control group, the cell proliferation, migration, and invasion abilities of the UHRF1 overexpression group were enhanced. After IWP-2 treatment, the cell proliferation, migration, and invasion abilities were inhibited. Knockdown experiment exhibited the reverse results to overexpression experiment. Conclusion UHRF1 may play an important role in the occurrence and development of CRC. UHRF1 overexpression may be a poor prognostic factor for CRC patients. UHRF1 may affect the proliferation, migration, and invasion of CRC cells through the WNT/MMP9 signaling pathway.

OBJECTIVETo investigate the effect of Kiss-1 gene suppression on the metastatic capacity of HCT116 human colorectal carcinoma cells in vitro and the involvement of nuclear factor-κB (NF-κB) signaling pathway.

METHODSA recombinant lentiviral vector of Kiss-1 gene pGC-LV-Kiss-1-EGFP or the empty vector was transfected in HCT116 cells. Cell Counting Kit-8 (CCK8) and Transwell chamber assay were used to detect the changes in cell proliferation, invasion and migration ability after the transfection. Western blotting was used to detect the expression of I-κB, the inhibitive protein of NF-κB signal pathway, and the expression of the downstream effector MMP-9 before and after transfection.

RESULTSIn cells over-expressing Kiss-1, I-κB expression increased and MMP-9 expression decreased significantly compared to those in the blank control and vector-transfected cells (P<0.05). Kiss-1 gene over-expression resulted in significant inhibition of HCT116 cell proliferation, invasion, and migration as compared to the control cells (P<0.05).

CONCLUSIONLentivirus-mediated Kiss-1 gene over-expression can inhibit the proliferation, invasion, and migration of HCT116 cells via the NF-B signaling pathway.

Cell Movement ; Cell Proliferation ; Colorectal Neoplasms ; pathology ; Genetic Vectors ; HCT116 Cells ; Humans ; I-kappa B Kinase ; metabolism ; Kisspeptins ; genetics ; Lentivirus ; Matrix Metalloproteinase 9 ; metabolism ; NF-kappa B ; metabolism ; Neoplasm Invasiveness ; Signal Transduction ; Transfection

Objective:To observe the efficacy and safety of teriprizumab combined with bevacizumab in above the second line treatment of high-level microsatellite instability (MSI-H) type metastatic colorectal cancer (mCRC) patients.Methods:From February 2019 to September 2019, 56 patients with MSI-H mCRC admitted to the Third Affiliated Hospital of Guangdong Medical University were selected and divided into control group and test group by random number table method, with 28 cases in each group. The control group was treated with bevacizumab, and the test group was treated with teriprizumab combined with bevacizumab. The objective response rate (ORR), disease control rate (DCR), progression-free survival, overall survival and incidence of adverse reactions were compared between the two groups.Results:The ORR and DCR of the test group were 60.71% (17/28) and 75.00% (21/28) respectively, higher than 28.57% (8/28) and 46.63% (13/28) of the control group, with statistically significant differences ( χ2=5.85, P=0.016; χ2=4.79, P=0.029). The median progression-free survival of patients in the control group and the test group were 3.5 months and 5.8 months respectively, with a statistically significant difference ( χ2=9.83, P=0.003). The median overall survival of patients in the control group and the test group were 12.1 months and 16.2 months respectively, with a statistically significant difference ( χ2=6.13, P=0.007). There were no significant diffe-rences in the incidences of hematological reaction (17.86% vs. 14.29%, χ2=0.13, P=0.716), cardiovascular injury (10.71% vs. 14.29%, χ2=0.16, P=0.686), liver and kidney function injury (25.00% vs. 21.43%, χ2=0.10, P=0.752), gastrointestinal reaction (28.57% vs. 35.71%, χ2=0.33, P=0.567), skin and mucosal injury (7.14% vs. 10.71%, χ2=0.35, P=0.553), nervous system disease (3.57% vs. 14.29%, χ2=2.25, P=0.134), endocrine reaction (3.57% vs. 10.71%, χ2=1.29, P=0.256), alopecia (14.29% vs. 17.86%, χ2=0.13, P=0.716) and fatigue (25.00% vs. 28.57%, χ2=0.27, P=0.605) between the control group and the test group. Conclusion:The combination of teriprizumab and bevacizumab can improve the short-term and medium-long-term efficacy of patients with MSI-H mCRC, which is safe and reliable.