Objective:To explore the regulation mechanism of the quorum sensing regulator AphA on the functional activity of type Ⅵ secretion system VflT6SS2 in Vibrio fluvialis. Methods:Western Blot analysis was used to detect the relative expression and secretion of VflT6SS2 signature component hemolysin-coregulated protein (Hcp) in wild type (WT), Δ aphA, and corresponding complementary strains. Quantitative reverse transcription PCR and luminescence activity assay of the promoter- lux fusion system was used to measure the mRNA expression levels and promoter activity of the VflT6SS2 core and accessory gene-cluster representative genes tssB2, hcp ( tssD2) and vgrG ( tssI2), and the quorum sensing regulator HapR in WT and Δ aphA strains. A point mutation experiment combined with a luminescence activity assay was used to verify the regulatory binding site of AphA in the tssD2b promoter region. Electrophoretic mobility shift assay (EMSA) was used to determine AphA binding to the hapR promoter. Results:The mRNA expression levels of tssB2, hcp( tssD2), vgrG ( tssI2), and hapR as well as the protein expression and secretion levels of Hcp in Δ aphA strain, were significantly higher than those in the WT strain. The promoter activities of the VflT6SS2 core cluster, tssD2a, tssI2a, and hapR were higher in Δ aphA strain than in the WT strain, while the promoter activity of tssD2b showed the opposite trend. The promoter sequence analysis of tssD2a and tssD2b found significant differences in the region from -335 bp to -229 bp, and two potential AphA binding sites on tssD2b. The promoter activity of tssD2b decreased significantly after the point mutation of the two potential AphA binding sites. EMSA results showed that AphA binds directly to the promoter region of hapR. Conclusions:AphA indirectly inhibits the regulation of the VflT6SS2 core and accessory gene clusters at the promoter level by directly repressing the expression of hapR. AphA showed opposite regulation patterns for tssD2a and tssD2b, and AphA could positively regulate the expression of tssD2b by directly binding to the tssD2b promoter region (-335 bp to -229 bp).

【Objective】 To explore the correlation between CSAG1 expression and the prognosis and tumor-infiltrating lymphocytes in renal clear cell carcinoma (RCCC), and to predict the survival and tumor progression. 【Methods】 The gene expression profiles and clinical information of CSAG1 were downloaded from the Cancer Genome Atlas (TCGA). Based on the differential mRNA expression, GO annotation and KEGG pathway analysis were performed. The relationship between CSAG1 and tumor immune infiltration was assessed with Tumor Immunoassay Resource (Timer 2.0) database. The mRNA expression of CSAG1 in human RCCC specimens was validated with qRT-PCR. 【Results】 CSAG1 expression was significantly higher in RCCC tissues than in normal tissues (P<0.05). The qRT-PCR results revealed that the mRNA level of CSAG1 was consistent with that predicted by bioinformatic analysis. The KEGG analysis and GO annotation indicated high GSAG1 expression in RCCC was related to transmembrane transport, tricarboxylic acid cycle and lysosome. CSAG1 expression was positively related to the infiltration of pDC, aDC, CD8⁺ T cells, cytotoxic cells, TFH, TH1 cells, Tem, NK CD56^dm cells, Treg and T cells, but negatively correlated with macrophage infiltration. 【Conclusion】 CSAG1 may be associated with poor prognosis of RCCC and become a potential immunotherapy target.

Objective: To explore the influence of parents educational level and family type on the level of parental control, so as to lay a foundation for improving parental education and promoting adolescents mental and physical health. Methods: In April 2019, random cluster sampling was used to select 1 046 students in junior and senior high schools in Bengbu. Information was collected through the Chinese version of Parental Control Questionnaire, and SPSS 25.0 for statistical analysis was used. Results: Mother s behavioral control and mother s educational level were significantly positively correlated( r =0.18), father s behavioral control and father s education level were significantly positively correlated( r =0.16). There was a significant positive correlation between the total score of maternal control and mother s education level( r =0.16). The total score of father s control was positively correlated with the education level of the fathers( r =0.12)( P <0.05). There was significant differences between the parents with different education level on parental control( H =16.93, 32.21, P <0.05). The score on parents behavioral control with primary school education or without education low among the parents with other education levels. There were differences between different family types in the total score of parental control( F =2.63, 3.55, P <0.05). Compared with extended families and nuclear families, single parent families total scored had significantly lower in father s control, and single parent families total scored significantly lower in mother s control scores than extended families. Parents educational level and family type had significantly predicted effect on parental behavioral control. Conclusion Parents educational level and family type have an influence on parents psychological and behavioral control, and have a predictive effect on parents control level. Parental control levels should be improved by improving the family s upbringing environment.

Objective: To explore the relationship between parental control and aggressive behavior of middle school students. Methods: Random cluster sampling was used to select 1 046 students.Information was collected through the Chinese version of Parental Control Questionnaire and Buss ＆ Perry Aggression Questionnaire. Results: The score on anger were high among female students and the total score were high among male students(t=6.28,2.56,P<0.01).Multiple comparison showed. the grade two junior middle school students’aggression level is significantly higher than grade three junior middle school students and grade one, two high school students(t=3.22,4.07,3.01,P<0.05). There were significant gender differences in parental control,father’s behavior control and father’s psychological control(P<0.05).Parental control shows a significant grade difference,multiple comparison showed, parental control among junior middle school students is significantly higher than that among high school students(t=3.22, 4.07, 3.01, P<0.05).Parental control and aggressiveness of the students were significantly positively correlated(r=0.16,0.29).There was a significant positive correlation between the psychological control of the parents and the aggressiveness of the boys(r=0.26,0.23).The behavior control and psychological control of the parents were positively correlated with the aggressiveness of the girls(r=0.09,0.13,0.29,0.35)(P<0.05).The Logistic regression analysis showed,father’s psychological control(OR=0.50,95%CI=0.21-0.79) and mother’s psychological control(OR=0.32,95%CI=0.02-0.62) showed significant positive association with aggression of the boys.Mother’s psychological control(OR=0.55,95%CI=0.23-0.87)and parents’ control(OR=0.17,95%CI=0.01-0.33) were positively associated with aggression of the girls(P<0.05). Conclusion Effective intervention of parental control might help prevent and control aggressive behavior of middle school students.

Objective: To understand the readiness for hospital discharge of the day surgery patients of obstructive sleep apnea-hypopnea syndrome and analyze its influencing factors. Methods: A total of 129 children with obstructive sleep apnea-hypopnea syndrome during the day surgery were investigated with a general data questionnaire and an adapted discharge preparation metric. Results: The total score of the readiness for hospital discharge was 166.38±30.93. The scores of discharge in all dimensions from high to low were adaptive ability 8.35±1.80, expected support 8.17±1.70, knowledge status 7.35±2.10, and personal status 7.10±1.43. Multiple linear regression results showed that the difficulty of caring for children from parents had a significant effect on the readiness of children with obstructive sleep apnea hypopnea syndrome during day surgery (P<0.01). Conclusions The discharge readiness of children with obstructive sleep apnea-hypopnea syndrome during day surgery was at the upper-middle level, but their personal status was not good. Medical and nursing staff should pay attention to health guidance for parents of children with disease-related knowledge, do a good job of interpretation before discharge, help parents increase their confidence in caring for children after discharge, improve the level of preparation for discharge, and ensure the safety of children after discharge.

In the process of kidney acquisition and transplantation, the donor kidney undergoes hypoxia, ischemia and reperfusion, which inevitably leads to ischemia/reperfusion (I/R) injury of the donor kidney. Hypoxia-inducible factor-1α(HIF-1α) is a ubiquitous oxygen-sensitive transcription factor. It can be activated rapidly during I/R injury. Then preventing and reducing the I/R injury of donor kidney during kidney transplantation by mediating the adaptation of donor kidney to hypoxic stress through direct or indirect ways, and participating in the repair of injury, thereby improving the effect of kidney transplantation. This review describes the evidence of the role of HIF-1α in I/R injury of donor kidney during kidney transplantation, and provides a theoretical basis for further research.

Objective To investigate the mechanism of renal injury induced by changes in flow shear stress (FSS) during renal ischemia/reperfusion (I/R).Methods 1.In vitro,HUVECs were divided into 4 groups:(1) HUVECs were loaded with 12 dyn/cm2 force for 30,45,and 90 min by using plate fluid chamber system.(2) Cells were loaded with FSS for 2 h,and then cultured for 1,3,8 and 12 h respectively;(3) HUVECs were pretreated with 0,1,2,4 and 8 mmol metformin and cultured for 24 h.(4) HUVECs in control group were cultured normally.The expression of p-AMPK/AMPK protein was detected by Western blotting in each group.2.In vivo,16 SD rats with successful establishment of IR model were randomly divided into 4 groups (n =4 in each group):(1) static cold storage (CS) group:isolated kidneys were stored for 4 h;(2) hypothermia machine perfusion (HMP) group:isolated kidneys were continuously perfused with 0 ℃ lactated Ringer's solution for 4 h;(3)metformin treatment group (Met-CS):metformin was intraperitoneally injected 3 days before surgery,and the isolated kidneys were obtained after cold preservation for 4 h;(4)rat kidneys of control group were just subjected to thermal ischemia for 30 min.The injury of renal tissue in each group was observed by TUNEL and HE staining.The expression and distribution of p-AMPK protein in renal tissues were detected by immunohistochemistry.The correlation between FSS loss and AMPK expression in kidney tissue was analyzed.Results The expression of p-AMPK in HUVECs could be up-regulated by FSS,and the expression of p-AMPK protein increased with the prolongation of time.After stopping FSS,the expression of p-AMPK protein in HUVECs gradually decreased with time (P＜0.05).Metformin could activate AMPK activity in a concentration-dependent manner (P＜0.05).The content of p-AMPK in renal tissue of HMP group was significantly higher than that of CS group (P＜0.05).The expression of p-AMPK in renal tissue of HMP group mainly distributed in the renal tubules,and few in glomerular endothelial cells and blood vessels.The apoptosis rate of renal tissue in HMP group was significantly lower than that in CS group (P＜0.05).In the HMP group,the damage of the renal tissue was mild,there was no swelling,and the renal tubules were slightly expanded.In the CS group,the renal tissue was severely damaged and the renal tubules were markedly swollen.Conclusion During the course of renal IR in rats,changes in FSS may affect renal tissue damage through the AMPK pathway.

Objective To investigate the relationship between thyroid hormone and uric acid (UA) and body mass index (BMI) in patients with type Ⅱ diabetes and normal thyroid function.Methods Total of 313 patients with type Ⅱ diabetes and normal thyroid function were selected.BMI,fasting blood glucose (FBG),the metabolism of blood lipid,thyroid hormones and UA indicators were examined and the correlations of thyroid hormone,BMI and UA were analyzed.Results (1) The patients were divided into two groups according to gender,and FT3,FT4,and UA of male were found to be significantly higher than those in female (P＜0.01).TSH,SBP,HDL-C in female were significantly higher than those in male (P＜0.01);(2) The patients were divided into three groups according to BMI Level.Thyroid-stimulating hormone (TSH),three iodine armour gland original glycine (TT3),free three iodine thyroid glycine (FT3),UA,and FBG in overweight and obesity groups were found to be higher than those in normal weight group (P ＜ 0.05);(3) The patients were divided into two groups according to the TSH level.Serum uric acid,TT3,FT3,fasting insulin in the group with TSH above 2.5 uIU/L were found to be higher than those in the group with TSH under 2.5 uIU/L (P ＜ 0.05);(4) Patients were divided into two groups according to the UA level.TSH,FT3 in male with high uric acid were found to be higher than those in male with normal uric acid (P ＜ 0.05);TSH was in female with high uric acid was found to be higher than that in female with normal uric acid (P ＜ 0.05).Conclusion Thyroid hormone in patients with type Ⅱ diabetes can be used to assess the body weight and uric acid,which is of great clinical importance.

Objective To observe clinical efficacy and explore clinical value of a modified procedure of double-stapling technique for mid-low rectal cancer.Methods Clinical data of patients undergoing laparoscopic anterior resection at the Department of General Surgery,the First Affiliated Hospital of Soochow University from February 2011 to February 2015 was analyzed retrospectively.According to the different ways in doing double-stapling technique,we divided patients into modified group (51 cases) and conventional group (74 cases).Parameters were compared between the two groups as general considerations,oncologic outcomes.Data were analyzed by SPSS 17.0 software packet,using t and x2 inspection.Results The difference of the general data of two groups was not statistically significant (P ＞ 0.05).Operation time in the modified group was longer than that of the conventional group [(169 ± 23) vs.(150 ±42)min,t =-3.150,P ＜0.05],but it had shorter drainage tube indwelling days [(7.9 ±2.9)d vs.(10.8±11.6)d,t=1.999,P＜0.05] and length of hospital stay after surgery [(10.0±3.6)d vs.(13.3 ± 13.7) d,t =1.025,P ＜ 0.05].The incidence of anastomotic leakage (2.0％ vs.18.9％,x2 =4.402,P ＜ 0.05) and tenesmus(3.9％ vs.17.6％,x2 =4.110,P ＜ 0.05) in the modified group was less than that of the conventional group.The difference in those areas was not statistically significant (P ＞ 0.05),such as intraopretive blood loss,per-anal exhaust time,consumption of liquid diet time,anastomotic bleeding,intestinal obstruction,reoperation for neostomy and infections.Conclusions Compared with traditional laparoscopic anterior resection,End-Corner anastomosis has the benefits of less postoperative anastomotic leakage and fewer low anterior resection syndrome.

OBJECTIVETo explore a new procedure of laparoscopic dual anastomosis for mid-low rectal cancer to reduce postoperative complications.

METHODSClinical data of 56 patients with mid-low rectal cancer undergoing laparoscopic rectal cancer resection(modified double-stapling technique, MDST, modification group) in the Department of General Surgery, the First Affiliated Hospital of Soochow University from February 2010 to June 2014 were compared with the data of 64 patients with mid-low rectal cancer (conventional double-stapling technique, DST, convention group) in the same period based on gender, age, tumor size, the distance from lower edge to the dentate line and tumor staging, etc. Patients in the modification group received operation as follows: (1) the rectum distal end was closed vertically instead of horizontally. (2) the anastomosis was conducted in an "end-corner" approach. (3) upper corner of the closed line in the distal end of rectum was removed. (4) the lower corner of closed line in the distal end of rectum was removed using vascular occlusion clamp method. (5) two T-shaped interchanges ("dangerous triangle") of stapled sutures formed after anastomosis were strengthened with absorbable suture. Patients in the convention group received laparoscopic dual anastomosis using conventional method: two corners and "dangerous triangles" were kept without any treatment. The clinical outcomes of two groups were analyzed retrospectively.

RESULTSThe intraoperational blood loss, postoperative drainage volume, postoperative anastomotic stoma bleeding, bowel function return and hospital stay were not significantly different between the two groups (all P>0.05). As compared to the convention group, the modification group had longer operation time [(211 ± 91) min vs. (174 ± 57) min, P<0.05], lower incidence of postoperative anastomotic leakage [1.8%(1/56) vs. 12.5% (8/64), P=0.030], lower tenesmus rate [3.6% (2/56) vs. 14.1% (9/64), P<0.05], less postoperative stoma re-creation [0 vs. 9.4% (6/64), P<0.05].

CONCLUSIONModified laparoscopic dual anastomosis for mid-low rectal cancer can significantly reduce the incidence of post-surgical complications such as anastomotic leakage.

Anastomosis, Surgical ; Anastomotic Leak ; Humans ; Laparoscopy ; Neoplasm Staging ; Operative Time ; Postoperative Complications ; Rectal Neoplasms ; surgery ; Retrospective Studies