Objective To investigate the role of purinergic 2X7 receptor (P2X7R) and its downstream target-NLRP3 inflammasome activation in the process of pancreatic fibrosis in a mouse model of chronic pancreatitis (CP). Methods Forty C57BL/6 mice were randomly divided into normal control group, CP group, P2X7R antagonist oxidized ATP (OxATP) group and brilliant blue G (BBG) group. The chronic pancreatitis model was induced by repeated intraperitoneal injection of the cholecystokinin analogue caerulein with the dose of 50μg/kg for six weeks. Normal saline, OxATP (20μL, 300μmol/L) or BBG (20μL, 10μmol/L) were administered for CP group, OxATP group and BBG group for two weeks after the last caerulein injection. Then all mice were sacrificed and the histopathological changes of the pancreas, especially the fibrotic degrees were evaluated by HE stain, fibrosis score, Sirius red staining and α-SMA immunohistochemical stain. The pancreatic P2X7R, NLRP3 and Caspase-1 expressions were detected by immunohistochemistry respectively to compare the changes between the groups, and explore the role of P2X7R-NLRP3 signaling pathway in pancreatic fibrosis. Results Compared with the normal control group, the scores of pancreatic fibrosis and the expressions of P2X7R, NLRP3 and Caspase-1 in pancreas were significantly increased in CP model group (P<0.05). Compare to CP group, the pancreatic chronic inflammation and the fibrosis indices such as HE fibrosis score, Sirius red staining and α-SMA immunohistochemical stain were ameliorated obviously in OxATP and BBG groups (P<0.05). And expressions of P2X7R, NLRP3 and Caspase-1 in the pancreas were allreduced greatly in both OxATP and BBG groups (P<0.05). Conclusion P2X7R antagonist OxATP and BBG can significantly decrease pancreatic chronic inflammation and fibrosis in the mouse model of CP, which suggests that the blockade of P 2X7R-NLRP3 inflammasome signaling pathway may represent a novel therapeutic strategy for CP and its fibrotic process.

Objective To explore the basic biological characteristics of lncRNA -uc.1 67,and its spatial dis-tribution,temporal expression pattern during the mouse embryonic development.Methods The UCSC genome browser of ENCODE was used to analyze preliminary bioinformatics of lncRNAs.Real -time (RT)-PCR was applied to detect the expression of uc.1 67 and neighboring genes in the embryonic mouse heart in different stages (P7.5,P1 1 .5,P1 4.5, P1 8.5).Dimethyl sulphoxide was used to induce P1 9 cell differentiation into the cardiomyocytes.RT -PCR was applied to detect the expression changes in uc.1 67 and neighboring genes on differential day 0,4,6,8 and 1 0.Results Full -length of human uc.167 was 201 bp,and human uc.167 was located in the genome 5q14.3 (chr5:88179623 -881 79824,GRCh37 /hg1 9).uc.1 67 mainly expressed in the ventricular muscle tissue.The expression of uc.1 67 was gradually decreased in the mouse embryonic heart development process(P7.5:1 .000 ±0.1 00,P1 1 .5:0.71 4 ±0.1 07, P1 4.5:0.393 ±0.043,P1 8.5:0.1 25 ±0.01 3),while the expression of its neighboring Mef2c gene was gradually in-creased(P7.5:1 .081 ±0.1 1 8,P1 1 .5:2.340 ±0.351 ,P1 4.5:3.958 ±0.542,P1 8.5:9.361 ±0.722),which showed an opposite trend.The expression of uc.1 67 during P1 9 cell differentiation into cardiomyocytes showed a an increase at first and then a decreasepattern,and the highest level expression of uc.1 67 was on differential day 4(d0:1 .071 ± 0.1 1 7,d4:4.71 4 ±0.501 ,d6:3.572 ±0.41 4,d8:2.550 ±0.31 4,d1 0:0.786 ±0.085).The expression of neigh-boring gene Mef2c was in an opposite trend(d0:1 .01 2 ±0.041 ,d4:0.353 ±0.037,d6:2.470 ±0.329,d8:6.706 ± 0.682,d1 0:7.765 ±0.705).Conclusions It is suggested that uc.1 67 may take part in the process of embryonic heart development and may play a role through negatively regulating its neighboring gene Mef2c.

OBJECTIVE:To study the effect and its mechanism of Fuzheng Jiedu Quyu formula(short forFuzheng formula) combined with oxaliplatin (L-OHP) on human colon cancer HT-29 cell proliferation and apoptosis. METHODS:HT-29 cells were divided into blank control group(without drugs),Fuzheng formula group(1000 mg/L),L-OHP group(31.25 mg/L)and combi-nation group (1000 mg/L Fuzheng formula+31.25 mg/L L-OHP). After cultured with corresponding drug for 48 h,MTT method was used to detect the cell proliferation;the changes of cellular morphology were observed by invert microscope;flow cytometry was used to detect the cell cycle and apoptosis rate. Proapoptotic gene Bax,apoptotic gene Bcl-2 mRNA expressions were deter-mined by real-time fluorescence quantitative polymerase chain reaction method;Bax,Bcl-2 protein expressions were assayed by Western blot. RESULTS:Compared with blank control group,cell proliferation was inhibited in L-OHP group and combination group;cell proportion was increased in S stage,G2/M stage and decreased in G0/G1 stage(P<0.05). Cell apoptosis rate in L-OHP group,Fuzheng formula group and combination group was increased;Bax mRNA and protein expression were up-regulated,Bcl-2 mRNA expression was downregulated(P<0.05);and combination group changed more obviously than the single drug groups(P<0.05). CONCLUSIONS:Fuzheng formula combined with L-OHP can inhibit HT-29 cell proliferation and promote its apoptosis, showing better effects than either of the two drugs alone. The mechanism may be associated with up-regulation of Bax gene and pro-tein expressions and down-regulation of Bcl-2 gene expressions in cells.

Objective:To express the glycoprotein D of herpes simplex virus type 2 (gD2) in the insect cells,and to determine its immunogenicity.Methods:HSV-2 genome was used as the template for amplification of gD2 extracellular domain fragment gene by PCR.The PCR product was inserted into the vector Bacmind,and the constructed recombinant plasmid gD2-Bacmind was transfected into the sf9 cells to package the recombinant baculovirus.The Sf9 cells were infected by recombinant baculovirus seed derived from the forth passage(P4),the titer of P4 recombinant baculovirus was detected by a plaque assay and the expression of recombinant protein gD2 was determined by Western blotting method.The supernatant of infected cells was collected and purified by Ni-NTA affinity chromatography to obtain the target protein gD2,the purified gD2 protein was used to immunize the BALB/c mice in 0, 2, 4 weeks (gD2 group),and PBS was used as negative control(PBS group);the titers of gD2 specific IgG in serum were detected by ELISA assay.Results: The PCR analysis and sequencing results proved that gD2-Bacmind was constructed correctly.The titer of recombinant baculovirus was 2.0×109 pfu·mL-1,the purified gD2 was about 37 000 with expectation,the percentage of gD2 in total protein was 90%.The average value of Log10 of titer of gD2 specific IgG in serum detected by ELISA assay in gD2 group at the sixth week was 4.34,and there was significant difference compared with PBS group(P<0.01).Conclusion: The gD2 expressed by insect-baculovirus expression vector system has the immunogenicity and can be selected as candidate protein for HSV-2 vaccine.

Objective:To investigate the association of tumor necrosis factor-α(TNF-α)gene G308A single nucleotide polymorphism(SNP)with childhood obesity and metabolic disorders.Methods:A total of 620 obese children admitted to our pediatric department from January 2015 to December 2019 were selected as research subjects, who were divided into metabolic non-healthy group( n=267)and metabolic healthy group( n=353), and 260 healthy children were selected as the control group. Single nucleotide polymorphism of TNF-α gene G308A was detected, serum TNF-α level, obesity indicators, glucose and lipid metabolism indicators were compared among the children in various groups. Results:No significant differences in body mass index(BMI), waist circumference, hip circumference, and serum TNF-α level were found between metabolic non-healthy group and metabolic healthy group( P>0.05), but higher than those in control group( P<0.05). There were no significant differences in fasting plasma glucose(FPG)and total cholesterol(TC) levels among the three groups( P>0.05). The levels of fasting insulin(FINS), fasting glucagon(FGC), HbA 1C, triglyceride(TG), and low density lipoprotein-cholesterol(LDL-C)in metabolic non-health group and metabolic health group were significantly higher than those in control group, higher in metabolic non-health group compared with metabolic healthy group( P<0.05). HDL-C level in two obese groups was significantly lower than that in control group, lower in metabolic non-health group compared with metabolic healthy group( P<0.05). The frequencies of GG genotype and allele G in metabolic non-healthy group and metabolic healthy group were significantly lower than those in control group, lower in metabolic non-health group compared with metabolic healthy group. The frequencies of AA, GA genotypes and allele A were significantly higher in metabolic non-health group than those in metabolic healthy group( P<0.05). There were no significant differences in TNF-α, FPG, and TC among AA, GA and GG groups( P>0.05). BMI, waist circumference, hip circumference, FINS, FGC, HbA 1C, TG, HDL-C and LDL-C were comparable between AA type and GA type( P>0.05), but revealed a significant difference compared with GG type( P<0.05). Pearson correlation analysis showed that the level of TNF-α was positively correlated with BMI, waist circumference, hip circumference, FPG, FINS, FGC, HbA 1C, TC, TG, and LDL-C( P<0.05), but negatively correlated with HDL-C( P<0.05). Conclusion:TNF-α gene G308A single nucleotide polymorphism is associated with obesity and metabolic disorders in children.

Objective Toinvestigatetheriskfactorsforleukoaraiosis(LA)inpatientswithlarge arteryatherosclerosis(LAA).Methods Theclinicaldata(age,sex,hypertension,diabetes,smoking, serum lipid level,hyperhomocysteinemia,and numbers of stenosis or occluded cerebral arteries)of 312 patients with LAA classified by the modified stop stroke study trial of Org 10172 in acute stroke treatment (SSS-TOAST ) were analyzed retrospectively. The age-related white matter changes (age related white matter changes,ARWMC)scale was used to evaluate LA. All the 312 patients were divided into non-LA group(n=72)and LA group(n=240)according the T2 weighted magnetic resonance imaging (MRI) and fluid attenuated inversion recovery(FLAIR)sequence,and 3 groups according to the (age-related white matter changes,ARWMC)scores:mild LA,moderate LA,and severe LA groups. The patients with multiple risk factors were analyzed by the univariate and multivariate Logistic regression analyses. Results (1)Of the 312 patients with LA,227 were males (72. 8%). Their average age was 64 ± 11 years,and 240 of them (76. 9%)had LA. Multivariate Logistic regression analysis showed that age (OR,2. 911,95%CI 1. 647-5.146,P=0. 000),hypertension (OR,2. 583,95%CI 1. 373-4.857,P<0. 01),diabetes (OR,1. 882, 95%CI 1. 058-3. 348,P <0. 05),the numbers of stenosis or occlusion arteries (OR,1. 851,95%CI 1.018-3. 367,P<0. 05),and lacunar infarction (LI)(OR,1.493,95%CI 1. 202-1. 853,P<0. 01)were the risk factors for LA. (2)The comparison of the clinical data in patients with different severity in the LA group found that there were significant differences in age,hypertension,diabetes,the numbers of stenosis or occlusionarteries,andLIamongthe3groups(allP<0.05).Conclusion Age,hypertension,diabetes, the numbers of stenosis or occlusion arteries,and LI are the independent risk factors for patients with LAA,and it is associated with the severity of LA.

Objective To investigate the relationship between platelet-activating factor acetylhydrolase gene Arg92His(4, 275; G→A), Ile198Thr(7, 593; T→C) and Val279Phe(9, 994; G→T) mutation and cerebral artery athero-sclerosis stenosis. Methods Six hundred forty-twopatients with cerebral infarction underwent cerebral digital subtrac-tion angiography (DSA).The patients were then divided into cerebral artery atherosclerosis stenosis (CAAS) group(n=477) and control group(n=81) accroding to the site and severity of their cerebral artery stenosis. Furthermore, the CAAS group were divided into intracranial artery stenosis(ICAS) subgroup(n=251), extracranial artery stenosis(ECAS) subgroup (n=115) and extracranial-intracerebral artery stenosis(ECAS) subgroup(n=111). The distributions of genotype and allele frequencies of Arg92His,Ile198Thr and Val279Phe mutation of platelet-activating factor acetylhydrolase gene were ex-amined and comparied in different groups. Results There were significant differences in the distributions of genotype and allele of Arg92His mutation between ICAS subgroup and control group(42.6% vs. 30.3%;23.3% vs. 16.4%, P <0.05). These associations were not detected in ECAS and IECAS subgroups. There was no significant association be-tween Ile198Thr and Val279Phe and stenosis at any site(P>0.05). The distributions of genotype and allele of Arg92His, Ile198Thr and Val279Phe mutation were no significantly difference between CAAS group and control group (P >0.05). Conclusions Arg92His mutation may be associated with intracranial artery atherosclerotic stenosis.

OBJECTIVETo explore the efficacy and safety of laparoscopic tunnel-like peri-anterior superior iliac spine suspension(L-TASISS) or combined with stapled transanal rectal resection (STARR) in the treatment of pelvic organ prolapse (POP) with outlet obstructive constipation (OOC).

METHODSA total of 119 POP patients complicated with OOC( II( to IIII( stage evaluated by POP-Q) received L-TASISS or combined with STARR in the First Affiliated Hospital of Zhengzhou University from August 2010 to January 2015. Clinical and follow-up data of these patients were analyzed retrospectively and compared before and after operation.

RESULTSAmong 119 cases, 51 cases(42.9%) underwent L-TASISS alone, and 68 cases (57.1%) received L-TASISS combined with STARR. All the 119 patients were successfully operated without conversion to open surgery. The operation time was (67.8±10.9) minutes, the intra-operative blood loss was (10.3±3.8) ml, the indwelling catheter time was (3.6±1.1) days, and hospital stay was (5.1±1.8) days. One month after operation, abdominal wall pain or stress occurred in 15 cases, of whom 3 cases were improved by local block injection, 1 case by incision stitches release, the rest ameliorated spontaneously within 3 months after surgery. No potential ureterostenosis, hydroureterosis, internal iliac vascular thrombosis resulting from compression of the mesh and other complications related to the mesh were found. One year after operation, all the patients were followed up. The OOC remission rate was 78.2%(93/119), of whom 76 cases were satisfied and 17 patients were completely satisfied. One case(0.8%) with stress urinary incontinence did not improve. Fifteen cases(12.6%) with algopareunia or dyspareunia did not achieve remission, but there was no new algopareunia or dyspareunia case. Eleven patients (9.2%) presented recurrence of symptoms, of whom 9 cases(7.6%) complained of sensation of incomplete evacuation. Two cases(1.7%) were graded as POP-Q II(, and 1 case (0.8%) as POP-Q III( after surgery. Constipation Score of all the patients was 1.4±0.9 (compared to 7.8±3.6 preoperatively) according to Rome III( criteria. Enterocele occurred in 53 cases (44.5%) preoperatively corresponded with only 1 case (0.8%) after operation (χ(2)=64.77, P=0.000). One hundred and six cases (86.6%) with defecation difficulties and 87 cases (73.1%) with sensation of incomplete evacuation before operation were significantly improved after surgery, corresponding with 7(5.9%) and 9 (7.6%) symptomatic cases respectively (χ(2)=155.78, P=0.000). Three cases (2.5%) with preoperative fecal incontinence symptoms were improved after operation as well.

CONCLUSIONThe procedure of L-TASISS or combined with STARR for POP patients with OCC has good short-term efficacy, and is safe and feasible.

Constipation ; Digestive System Surgical Procedures ; methods ; Fecal Incontinence ; Humans ; Laparoscopy ; Middle Aged ; Operative Time ; Pelvic Organ Prolapse ; surgery ; Rectal Prolapse ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate the characteristics of gastrointestinal transit in chronic constipation patients depending on a small amount barium(SAB) gastrointestinal transit test.

METHODSImaging data of 1890 cases with chronic constipation diagnosed with Rome III( criteria undergoing the SAB gastrointestinal transit test at Department of Colorectal Surgery, The First Affiliated Hospital of Zhengzhou University from January 2009 to March 2015 were retrospectively analyzed. All the patients took 20 g medical barium sulfate diluted in porridge and erect position abdominal X-ray photographs were then taken at 4, 8, 12, 24 hours after meal and once in every 24 hour sequentially. Transit characteristics of whole gut and various segments, including stomach, small intestine, right hemicolon, left hemicolon and rectum were assessed according to the predetermined criteria.

RESULTSAll the 1890 patients showed good compliance with the complete test. Among these patients, 794 cases(42%) were diagnosed as normal transit and 1096 cases(58%) as gastrointestinal slow transit. Classified by the site of slow transit, 151 cases(8%) were abnormal gastric transit, 175(9%) cases were abnormal small intestinal transit, 986(52%) cases were slow right hemicolon transit, 1039 cases(55%) were slow left hemicolon transit, 139 cases(7%) were outlet obstruction. Among the 1096 patients with slow gut transit, 907 cases(83%) were multiple segments slow transit and 189 cases(17%) were single segment slow transit. Among 907 patients with multiple segmental slow transit, 668 cases(74%) were total colon slow transit, 61 cases(7%) were colonic slow transit combined with outlet obstruction, 138 cases(15%) were small intestine slow transit coincided with colon slow transit, 40 cases(4%) were total segments slow transit in the whole gut. Of 189 cases of single segment slow transit, 17 cases(9%) were unique gastric delayed empting, 37 cases(20%) were unitary small intestine transit dysfunction, 19 cases(10%) were right hemicolon transit defect, 78 cases(41%) were left hemicolon transit deterioration, 38 cases(20%) were outlet obstruction.

CONCLUSIONSThe characteristics of gastrointestinal transit test in patients with chronic constipation varied from each other. Majority presents multi-segment slow transit combined with colon slow transit. SAB gastrointestinal transit test is helpful for surveying the transit characteristics of each segment of gut and worth clinical generalization and application.

Colon ; Constipation ; Gastrointestinal Tract ; Gastrointestinal Transit ; Humans ; Rectum ; Retrospective Studies ; Stomach

The curative efficacy of percutaneous transluminal angioplasty and stenting (PTAS) in the treatment of patients with ischemia cerebrovascular disease caused by artery stenosis was explored. The clinical data of 111 patients with ischemia cerebrovascular disease receiving PTAS in Guangdong Province General Hospital from Aug. 2007 to Nov. 2009 were retrospectively analyzed. In total 132 stents were implanted in the 111 patients. The mortality and rate of neural and non-neural complications were assessed perioperatively. Outcomes [including the frequency of transient ischemic attack (TIA), stroke, or death from vascular diseases) were assessed after operation. NIHSS rating was performed in all cases before and at first week, 6th month and 12th month after the operation. The PTAS success rate was 100%. The degree of stenosis was reduced after PTAS. The total complication rate during perioperative period was 15.3% (the rate of neural complications was 3.6%). Sixty-seven patients were followed up. Three patients (4.48%) developed cerebrovascular events within 1 month, containing one case of TIA, one case of ipsilateral mild stroke and one case of contralateral mild stroke. No severe stroke or death was observed. During a follow-up period of 12 months 7 patients had cerebrovascular events (10.44%), including 2 cases of ipsilateral TIA (2.99%), 2 cases of ipsilateral mild stroke and 2 cases of contralateral mild stroke (2.99%), one case of severe stroke (1.49%). In 13 patients receiving DSA re-examination one year after PTAS, 2 patients (15.38%) had in-stent restenosis. NIHSS scores were obviously decreased during a follow-up period as compared with those pre-operation (P<0.05). It was concluded that PTAS could significantly alleviate the neural function deficit of the patients with ischemia cerebrovascular disease. The success rate of PTAS was high, and the rate of complications was lower and the clinical outcomes were satisfactory. PTAS is a safe and effective therapeutic method, though the long-term outcomes need further study.