ABSTRACT
Objective:To explore the expression and clinical significance of Janus protein tyrosine kinase/signal transducer and activator of transcription (JAK3/STAT5) signaling pathway in peripheral blood mononuclear cells (PBMCs) of patients with primary gouty arthritis (GA).Methods:Peripheral blood, clinical data and laboratory tests were collected from 50 patients with acute gout (AG), 50 patients with intermittent gout (IG) and 50 healthy controls (HC). Quantitative real-time-polymerase chain reaction (RT-qPCR) was used to detect mRNA expression level of JAK3/STAT5 related genes (JAK3, STAT5a, STAT5b). Enzyme linked immune sorbent assay (ELISA) was used to detect interleukin-2 (IL-2) concentration in subject′s plasma. Measurement data among the three groups that was in accordance with normal distribution was analyzed by one-way analysis of variance, pairwise comparisons using LSD, non-normal distribution data was analyzed by Mann-Whitney test or Kruskal-Wallis H test, and correlation analysis between variables was analyzed using Spearman correlation analysis. Results:① The mRNA expression levels of JAK3, STAT5a and STAT5b were significantly different among the three groups ( F=50.13, P<0.01; F=7.573, P=0.000 7; F=12.14, P<0.01), of which the JAK3 mRNA expression level in the HC group [(606±65)×10 -4] was significantly higher than that in the AG group [(103±13)×10 -4] and IG group [(114±24)×10 -4], and the difference was statistically significant (both P<0.01), while the STAT5a mRNA expression level in the AG group [(89±9)×10 -4] was significantly higher than that in the IG group [(59±4)×10 -4] and HC group [(61±4)×10 -4], and the difference was statistically significant ( P=0.002, P=0.003 9), and the expression level of STAT5b mRNA in HC group [(60±5)×10 -4] was significantly lower than that in AG group [(95±7)×10 -4] and IG group [(98±7)×10 -4], and the difference was statistically significant ( P=0.000 2, P<0.000 1). ② The difference of IL-2 concentration in plasma among the three groups was statistically significant ( F=22.87, P<0.01), and the serum IL-2 concentration in the AG group [(87.9±8.4) pg/ml] was significantly higher than that in the IG group [(32±4) pg/ml] and HC group [(44±4) pg/ml], and the difference was statistically significant (both P<0.01). ③ Spearman correlation analysis showed that the mRNA expression of STAT5a and STAT5b was positively correlated with the absolute value of neutrophils in patients with gout ( r=0.282, P<0.05; r=0.257, P<0.05). Conclusion:The IL-2/JAK3/STAT5 signaling pathway is involved in the occurrence and development of gout, suggesting that this pathway may play a key role in the pathogenesis of gout.
ABSTRACT
Objective:To explore the expression and clinical significance of late autophagy in per-ipheral blood mononuclear cells (PBMCs) of patients with primary gouty arthritis (GA).Methods:Peripheral blood, clinical data, and laboratory tests were collected from 30 patients with acute gout (AG), 30 patients with intermittent gout (IG), and 50 healthy controls (HC). Quantitative polymerase chain reaction (RT-qPCR) was used to detect mRNA expression levels of autophagy-related genes (ATG5, ATG12, ATG16, ATG3, ATG7, ATG10, ATG4B, LC3-2/LC3B). Measurement data conformed to normal distribution were tested using t test or analysis of variance (ANOVA), and non-normal distribution data were tested using Mann-Whitney test or Kruskal-Wallis H test. SNK was used for pairwise comparison among the three groups. Correlation between variables was tested by Spearman correlation analysis. Results:① The expression level of ATG5 mRNA,ATG12 mRNA, ATG16 mRNA, ATG10 mRNA and LC3-2 mRNA in the AG group was lower than that of the IG group and the HC group, and the expression level of the IG group was lower than that of the HC group[9.16×10 -3(6.04×10 -3, 15.00×10 -3) vs 14.48×10 -3(9.95×10 -3, 21.38×10 -3) vs 0.08×10 -3(12.21×10 -3, 42.79×10 -3), H=19.377, P<0.001; 18.89×10 -3(13.85×10 -3, 24.92×10 -3) vs 21.13×10 -3(12.11×10 -3, 28.06×10 -3) vs 33.57×10 -3(13.11×10 -3, 49.89×10 -3), H=7.545, P=0.023; 8.72×10 -3(4.96×10 -3, 13.74×10 -3) vs 10.62×10 -3(7.48×10 -3, 24.71×10 -3) vs 20.07×10 -3(11.99×10 -3, 39.56×10 -3), H=20.962, P<0.001; 1.05×10 -3(0.73×10 -3, 1.84×10 -3) vs 1.60×10 -3(0.93×10 -3, 2.58×10 -3) vs 1.69×10 -3(1.05×10 -3, 3.54×10 -3), H=8.193, P=0.017; 2.31×10 -3(1.22×10 -3, 3.53×10 -3) vs 2.78×10 -3(1.68×10 -3, 5.96×10 -3) vs 3.68×10 -3(2.00×10 -3, 5.67×10 -3) , H=7.135, P=0.028]. The expression level of ATG4B mRNA in the AG and IG group was higher than that in HC group, and there was significant difference between IG group and AG group, IG group and HC group[9.95×10 -3(6.32×10 -3, 12.23×10 -3) vs 10.86×10 -3 (8.80×10 -3, 17.03×10 -3) vs 8.07×10 -3(5.52×10 -3, 11.63×10 -3), H=8.531, P=0.014]. There was no significant difference between the ATG3 mRNA and ATG7 mRNA groups ( H=0.539, 3.739, bothall P values >0.05). ② The results of Spearman correlation analysis suggested that in patients with acute gout, ATG3 was negatively correlated with PDW and MPV ( r=-0.499, P=0.006; r=-0.463, P=0.011); ATG4B was positively correlated with HDL-C ( r=0.408, P=0.048); ATG7 was negatively correlated with GLOB ( r=-0.554, P=0.001); ATG10 was positively correlated with ALB ( r=0.412, P=0.024) and negatively correlated with Crea and hsCRP ( r=-0.459, P=0.011; r=-0.375, P=0.045); ATG12 was negatively correlated with MO ( r=-0.434, P=0.017); ATG16 was negatively correlated with ALT and AST ( r=-0.389, P=0.034; r=-0.366, P=0.047); LC3-2 was positively correlated with UA ( r=0.381, P=0.041) and negatively correlated with MPV and PDW ( r=-0.413, P=0.026; r=-0.449, P=0.015). In patients with intermittent gout, ATG3 and ATG4B were negatively correlated with apoB100 ( r=-0.555, P=0.011; r=-0.462, P=0.040); ATG5 was negatively correlated with Crea ( r=-0.456, P=0.011); ATG10 was negatively correlated with TC, LDL-C, and apoB100 ( r=-0.526, P=0.017; r=-0.556, P=0.011; r=-0.515, P=0.020). Conclusion:Autophagy is involved in the development of gout, and is correlated with ibflammatory and metabolic indicators, suggesting that autophagy is an important feature in the pathogenesis of GA.
ABSTRACT
Objective:To investigate the possible role of miR-146a in the patho-genesis of inflammation in primary gout arthritis.Methods:① The RAW264.7 mouse macrophage was stimulated with 200 μg/ml monosodium urate (MSU) crystal for 0 h, 3 h, 6 h, and 12 h. Then cells and super-natants were collected. The miR-146a was detected by TaqMan probe method. The expression of interleukin-1 receptor-associated kinase 1 (IRAK 1), tumor necrosis factor receptor-associated factor 6 (TRAF6), nuclear factor-kappa B (NF-κB), interleukin (IL)-1β, tumor necrosis factor-α (TNF-α) mRNA were detected by real-time (RT)-quantitative polymerase chain reaction (qPCR). The concentration of IL-1β was measured in the culture supernatant by enzyme-linked immunosorbent assay (ELISA). The protein expression levels of TRAF6, NF-κB and IL-1β were detected by Western-blotting. ② The RAW264.7 mouse macrophage was transfected with miR-146a mimics, miR-146a mimic control, miR-146a inhibitor, and miR-146a inhibitor control. After stimulating each group of cells with 200 μg/ml MSU crystals for 6 h, the expression of miR-146a, IRAK1, TRAF6, NF-κB, IL-1β mRNA and TRAF6, NF-κB, IL-1β protein were measured. The measurement data were compared by Independent sample t test. and repeated measures analysis of variance (ANOVA). Results:① After MSU crystals stimulated RAW264.7 cells, we found that the expression level of miR-146a in the stimulation group at 3 h, 6 h, and 12 h was lower than that in the control group ( t=-10.234, -17.059, -26.204, P<0.01), and then, IL-1β protein concentration at 6 h, 12 h was higher ( t=7.552, 9.007, P<0.01). Meanwhile, IRAK1, TRAF6, NF-κB and IL-1β mRNA in the stimulation group at 3 h and 6 h were higher than those in the control group ( t=9.847, 6.147, P<0.01; t=3.49, 3.32, P<0.05; t=3.643, 8.471, P<0.05; t=8.726, 49.68, P<0.01). TNF-α mRNA at three time points in the stimulation group was high ( t=4.691, 11.115, 12.816, P<0.01). Moreover, the results showed that the relative expression of TRAF6 and NF-κB protein in the stimulation group at 6 h and 12 h was higher than that in the control group ( t=8.052, 8.119, P<0.01, t=22.454, 5.845, P<0.01), IL-1β protein in the stimulation group increased at all three time points compared with the control group ( t=18.561, 4.74, 8.432, P<0.01). ② After trans fection, the miR-146a mRNA expression of the mimics group was significantly higher than the mimics control group ( t=31.769, P<0.01); the inhibitor group was significantly lower than the inhibitor control group ( t=-4.22, P<0.05). ③miR -146a overexpression group was stimulated with 200 μg/ml MSU crystals for6 h, the expression levels of IRAK 1, TRAF 6, NF-κB and IL-1β mRNA in the mimic group were lower than those in the mimic control group ( t=-14.754, -21.201, -19.381, -17.323, P<0.01), the expression levels of TRAF 6, NF-κB and IL-1β protein were also lower than those in the mimic control group ( t=-3.137, -32.974, -18.789, P<0.05), while the inhibitor group had good results. Conclusion:① Overexpression of miR-146a can reduce the expression of IRAK1, TRAF6, NF-κB, IL-1β and inhibit MSU crystal-mediated inflammation, while inhibition of miR-146a expression can aggravate inflammation, suggesting that miR-146a participates in the negative feedback regulation of gout inflammation. ② miR-146a may target the NF-κB signaling pathway and participate in spontaneous remission of gouty arthritis.
ABSTRACT
Objective To investigate the safety and feasibility of the modified terminal cannula ileostomy in laparoscopic anus-preserving operation of low rectal cancer (RC).Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 15 patients who underwent laparoscopic radical resection of RC + terminal cannula ileostomy in the Subei People's Hospital of Jiangsu Province between September 2016 and June 2017 were collected.The patients underwent laparoscopic low anterior resection of RC,intra-abdominal sigmoid colon-rectum end-to-end anastomosis after extracting tumor specimens,and terminal cannula ileostomy in vitro.Observation indicators:(1) intraoperative situations:operation time,time of cannula ileostomy,volume of intraoperative blood loss,number of lymph node dissected,surgical margin;(2) postoperative situations:time to initial anal exsufflation,recovery time of defecation,time of tube removal,closing time of stoma,postoperative complications,duration of hospital stay;(3) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect the anastomotic leakage-related complications up to December 2017.Measurement data with normal distribution were represented as (x)±s.Results (1) Intraoperative situations:all the 15 patients underwent successful laparoscopic radical resection of RC + terminal cannula ileostomy,without conversion to open surgery and death.The operation time,time of cannula ileostomy,volume of intraoperative blood loss and number of lymph node dissected were respectively (170 ± 34) minutes,(23 ± 4) minutes,(59 ± 27)mL and (13 ± 5) per case.No residual cancer cells were found in resection margins.(2) Postoperative situations:time to initial anal exsufflation and recovery time of defecation in 15 patients were respectively (6± 1) days and (7 ± 1) days.The stoma was automatically closed after tube removal,and time of tube removal and closing time of stoma after tube removal were respectively (23 ± 2) days and (3 ± 1) days.The incidence of postoperative complications was 1/15.One patient with catheterization-related complications was improved by strengthening dressing and antibiotic use,and the stoma was healed at 8 days after tube removal.There was no anastomotic leakage-related complications and death.The duration of hospital stay was (15±3) days.(3) Followup situations:all the 15 patients were followed up for 6-12 months.During the follow-up,there were no anastomotic leakage-related complications and death.Conclusion The modifyied terminal cannula ileostomy is safe and feasible,and is also an ideal surgical method for preventing anastomotic leakage in the laparoscopic anuspreserving operation of low RC.
ABSTRACT
Objective To explore the effects of herceptin on the serum tumor markers in patients with HER-2 positive advanced breast cancer treated with chemotherapy and its influence on the immune function.Methods Seventy-six patients with HER2-positive advanced breast cancer from January 2013 to June 2014 in Xiangyang No.1 People's Hospital Affiliated to Hubei University of Medicine were divided into the observation group (38 cases) and the control group (38 cases),fifty healthy subjects in the same period were selected as the blank group.The control group received gemcitabine combined with capecitabine,gemcitabine 1000 mg / m2 via intravenous infusion in the first and eighth day,capecitabine 2500 mg/m2 via intravenous drip from the first day to fourteenth day;the observation group were given trastuzumab on the basis of the control group,4 mg/kg for the first time,then 2 mg/kg,1 time/week,three weeks for a course,have been given for four courses.The peripheral blood lymphocyte subsets were detected by flow cytometry,the Carcinoembryonic antigen (CEA),cancer-associated glycoprotein antigen (CA153) and tissue polypeptide specific antigen (TPS) were detected by enzyme-linked immunoassay.The short-term efficacy,survival and adverse reactions incidence rate in the two groups were compared.Results (1) The clinical benefit rate of the observation group was higher than that of the control group (89.47% vs.63.15%,χ2=8.508,P0.05),the CD3+,CD4+,CD4+/CD8+ of the observation group and the control group after treatment were significantly lower than those of the blank group (F=14.484,9.371,8.213,P<0.01).(3) The CEA,CA153 and TPS of the observation group and the control group after treatment were (5.05±2.03) μg/L,(7.25±2.70)μg/L,(36.04±12.05) IU/ml,(55.42±18.23) IU/ml,(262.43±53.38) IU/ml,(355.21±47.80) IU/ml,which were significantly lower than those before treatment (t=8.743,3.805,7.929,5.271,9.512,6.389,P<0.05),the observation group was significantly lower than the control group (t=3.353、3.665、3.442,P<0.05).(4) The incidence rate of pain and fever in the observation group were 34.21% and 28.95%,respectively,which were significantly higher than those in the control group,the difference was statistically significant (χ2=15.683,12.862,P<0.01).Conclusion Trastuzumab can synergistically reduce the serum tumor marker content in the treatment of patients with HER 2-positive advanced breast cancer,effectively improve the short-term efficacy of conventional chemotherapy,extend the life span,without the cause of significant damage to the immune function,but it can significantly increase the incidence rate of pain and fever,so it is necessary to carry out targeted method to improve chemotherapy compliance.
ABSTRACT
Objective To investigate the safety and feasibility of the reversed π-shaped esophagojejunal anastomosis in laparoscope-assisted total gastrectomy (LATG) for gastric cancer (GC).Methods The retrospective corss-sectional study was conducted.The clinicopathological data of 18 GC patients who were admitted to the Subei People's Hospital of Jiangsu Province between January 2015 and October 2016 were collected.All the 18 GC patients underwent LATG,surgical procedures included free stomach and lymph node dissection firstly,side-to-side jejunal anastomosis secondly and laparoscopic gastrointestinal reconstruction using reversed π-shaped anastomosis finally.Observation indicators:(1) surgical situations:side-to-side jejunal anastomosis method,conversion to open surgery,operation time,reversed π-shaped anastomosis time,volume of intraoperative blood loss and number of lymph node dissected;(2) postoperative situations:time for initial out-of-bed activity,time to initial anal exsufflation,time for postoperative water intake,time of drainage tube removal,postoperative complications and duration of hospital stay;(3) postoperative pathological examination;(4) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect diet intake,anastomosis patency,gastrointestinal obstruction and patients' survival up to March 2017.Measurement data with normal distribution were represented as (x)±s.Results (1) Surgical situations:18 patients underwent successful LATG and reversed π-shaped esophagojejunal anastomosis,without conversion to open surgery and perioperative death.Five patients used in vitro hand-sewn side-to-side esophagojejunal anastomosis through small incision of specimens sampling,and 13 completed all surgery under laparoscopy.Operation time,reversed π-shaped anastomosis time,volume of intraoperative blood loss and number of lymph node dissected of 18 patients were (187±12)minutes,(37±5) minutes,(735±18)mL and 29±2,respectively.(2) Postoperative situations:time for initial out-of-bed activity,time to initial anal exsufflation,time for postoperative water intake and time of drainage tube removal in 18 patients were (1.6±0.5) days,(2.3±0.4) days,(2.5±0.5) days and (7.5± 1.5) days,respectively.One patient complicated with esophagojejunal anastomosis fistula at postoperative day 3 was cured by drainage and symptomatic treatment and then discharged from hospital.Eighteen patients received regularly angiography using oral water-soluble contrast medium after recovering fluid diet intake,showing anastomosis patency and no contrast medium leakage,and then discharged from hospital.Duration of hospital stay of 18 patients was (12± 11) days.(3) Postoperative pathological examination:of 18 patients,15 were diagnosed with adenocarcinoma and 3 with signet-ring cell carcinoma.T2,T3 and T4 of T staging were respectively detected in 3,3 and 12 patients.N0,N1,N2 and N3 of N staging were respectively detected in 8,3,2 and 5 patients.Stage Ⅰ,Ⅱ and Ⅲ of TNM stage were detected in 3,5 and 10 patients,respectively.(4) Follow-up and survival situations:17 of 18 patients were followed up for 6-25 months,with a median time of 12 months.During the follow-up,2 patients were complicated with sour regurgitation and vomiting after eating at month 6 and 12 postoperatively and received gastrointestinal contrast examination,showing anastomotic stenosis,and then were cured by endoscopic dilation and discharged form hospital.Other patients had good diet and survival,without anastomotic complications.Conclusion The reversed π-shaped esophagojejunal anastomosis in LATG for GC is safe and feasible,with good short-term outcomes.
ABSTRACT
Digestive tract reconstruction is an important part of gastric carcinoma operation as well as tumor resection and lymph node dissection.Surgeons are seeking the optimal reconstruction method that reduces the occurrence of complications and maintains better quality of postoperative life extremely.Uncut Roux-en-Y anastomosis is a modified procedure in which an artificial jejunal occlusion is devised to avoid Roux stasis syndrome based on Billroth Ⅱ and Braun's anastomosis.Compared to the conventional Roux-en-Y anastomosis,the uncut Roux-en-Y anastomosis retains the advantage of preventing biliary and pancreatic secretions reflux,furthermore,it can decrease the symptoms associated with Roux stasis owing to the abnormal myoelectrical conduction of Roux limb.Because the early studies indicated that there was higher incidence of dehiscence or recanalization of the jejunal occlusion,the uncut Roux-en-Y anastomosis has not been widely applied.Since jejunal occlusion has been optimized recently,the uncut Roux-en-Y reconstruction may be an optional and appropriate method of digestive tract reconstruction after distal gastrectomy.
ABSTRACT
Objective To investigate the application and clinical effect of uncut Roux-en-Y (uncut RY) anastomosis in laparoscopic distal radical gastrectomy of gastric cancer.Methods The retrospective crosssectional study was adopted.The clinical data of 23 patients with gastric cancer who were admitted to the Northern Jiangsu People's Hospital from December 2014 to July 2015 were collected.All the 23 patients underwent laparoscopy-assisted distal gastrectomy (LADG) and total laparoscopic distal gastrectomy (TLDG) according to the individual situations.The indexes of observation were collected,including (1) intraoperative indexes:operation time,uncut RY anastomosis time and volume of inraoperative blood loss,(2) postoperative indexes:time to anal exsufflation,time for initial water intake,time for semi-fluid diet intake,time for out-off-bed activity,duration of hospital stay,occurrence of complications and results of pathological examination,(3) results of follow-up.The follow-up was performed by outpatient examination and telephone interview up to November 2015,including postoperative discomfort after diet intake,barium meal examination of gastrointestinal tract at postoperative month 1 (anas-tomotic stenosis,recanalization and dehiscence of occlusion),detecting situations of gastric remnant and anas-tomotic stoma at postoperative month 3 by gastroscopy and occurrence of gastrointestinal obstruction.Measurement data with normal distribution were presented as x ± s.Results (1) Intraoperative situations:all the 23 patients underwent successful uncut RY anastomosis,including 18 receiving LADG and 5 receiving TLDG.The operation time,uncut RY anastomosis time and volume of intraoperative blood loss were (165.9 ± 11.6) minutes,(18.2 ± 2.2) minutes,(48 ± 6) mL in all the 23 patients and (172.0 ± 8.5) minutes,(26.6 ± 1.5) minutes,(46 ± 4) mL in 5 patients with TLDG,respectively.Two patients received hemostatic treatment using suture and hemostatic forceps due to anastomotic bleeding.(2) Postoperative situations:time to anal exsufflation,time for initial water intake,time for semi-fluid diet intake,time for out-off-bed activity,duration of hospital stay and incidence of complications in all the 23 patients were (2.2 ± 0.4) days,(2.7 ± 0.4) days,(3.5 ± 0.4) days,(2.7 ± 0.3) days,(10.6 ± 1.4) days and 8.7％ (2/23),respectively.No patient was dead in the perioperative period.Two patients complicated with incisional infection and high fever were cured by symptomatic treatment,without occurrence of anastomotic leakage,bleeding and anastomotic-related complications.All the patients received postoperative barium meal examination of upper gastrointestinal tract,with unblocked anastomotic stoma and without leakage of barium meal.Diameter of tumor and number of lymph node dissected were (3.2 ± 1.2) cm and 30 ± 4,with negative upper and lower resection margins.Numbers of patients with tumor differentiation,T stage,N stage and TNM stage were 12 and 11 in differentiated and undifferentiated tumors,1,9 and 13 in T1,T2 and T3 stages,9,11 and 3 in N0,N1and N2 stages,1,4,9,6 and 3 in Ⅰ a,Ⅰ b,Ⅱ,Ⅲ a and Ⅲ b stages,respectively.(3) All the 23 patients were followed up by outpatient examination for 3-11 months.One patient had discomfort in upper abdomen with vomiting at postoperative week 3,and no anastomotic leakage,bleeding and anastomotic-related complications were occurred in other patients.Conclusion As a modified anastomotic method,uncut RY anastomosis is safe and feasible,and it is also an ideal method of digestive tract reconstruction after laparoscopic distal radical gastrectomy.
ABSTRACT
In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor (LIF) in mice with embryonic implantation dysfunction (EID), 120 Kunming mice post coition were randomized into three groups: normal control group, model group and traditional Chinese medicine group (TCM group) (n=40 in each group). Uterus was collected on the pregnancy day (Pd) 4, 5, 6 after an intravenous injection of Evan's blue. The endometrium was dyed by Evan's blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of LIF mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve embryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.