Effects of Helicobacter pylori infection on m6 A methylation level and related enzymes expression in gastric epithelial cells / 安徽医科大学学报

Objective @# To investigate the expression of N6-methyladenosine modification ( m6 A) and related en- zymes FTO，METTL3 and YTHDF2 in gastric epithelial cells infected with Helicobacter pylori (Hp) ，and to analyze its expression and clinical significance in gastric cancer by using online websites．@*Methods @#Gastric epithelial cells GES-1 and AGS were infected with Hp GZ7 strain，and the multiplicity of infection was 30 ∶ 1．Total RNA and total protein were collected after infecting 24 h，and the m6 A modification level of total RNA was detected by RNA dot hybridization．The mRNA and protein expressions of FTO ，METTL3 and YTHDF2 were detected by q-PCR and Western blot．The expression difference of FTO，METTL3 and YTHDF2 in gastric cancer tissues and normal tissues and their expression in Stage classification were analyzed by GEPIA，and the relationship between the expression of FTO，METTL3 and YTHDF2 and prognosis was analyzed by Kaplan-Meier Plotter. @*Results @# Compared with the un- infected group，the m6 A modification level of total RNA in both Hp-infected GES-1 and AGS cells significantly de- creased，the mRNA and protein expression levels of FTO increased，and the mRNA and protein levels of METTL3 and YTHDF2 decreased，with statistical significance (P＜0. 05) ．The expression level of FTO in gastric cancer tis- sues was higher than that in normal tissues，with statistical significance (P＜0. 05) ，YTHDF2 was higher than that in normal tissues，with no statistical significance，and the expression level of METTL3 was lower than that in nor- mal tissues，with no statistical significance．Patients with high expression of FTO and METTL3 had a lower survival rate，while patients with high expression of YTHDF2 had a higher survival rate．The expression of FTO was signifi- cantly correlated with Stage classification (P＜0. 05) ，while the expression of METTL3 and YTHDF2 was not sig- nificantly correlated with Stage classification．@*Conclusion @# Hp infection can change the level of m6 A and the ex- pression of related enzymes FTO，YTHDF2 and METTL3，suggesting that m6 A methylation may play a role in the occurrence and development of Hp-induced gastric cancer.

Role of Nod-like receptor family pyrin domain-containing 2 in dorsal root ganglion in neuropathic pain in rats / 中华麻醉学杂志

Objective:To evaluate the role of Nod-like receptor family pyrin domain-containing 2 (NLRP2) in the dorsal root ganglion (DRG) in neuropathic pain (NP) in rats.Methods:Thirty-two male Sprague-Dawley rats in which intrathecal catheters were successfully implanted, aged 2-3 months, weighing 200-250 g, were divided into 4 groups ( n=8 each) using a random number table method: sham operation group (S group), NP group, NP plus NLRP2-siRNA group (NP+ siRNA group) and NP plus NLRP2-scrRNA group (NP+ scrRNA group). The right sciatic nerve was only exposed but not ligated in group S. NP was induced by chronic constriction injury (CCI) to the sciatic nerve in anesthetized rats in group NP, group NP+ siRNA and group NP+ scrRNA.NLRP2-siRNA and NLRP2-scrRNA were intrathecally injected at 3 days before CCI in group NP+ siRNA and group NP+ scrRNA, respectively.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before CCI (T 0) and 1, 3, 7 and 10 days after CCI (T 1-4). The rats were sacrificed after the last measurement of pain threshold, and the L 4, 5 segments of the DRG on the operated side were removed for determination of the expression of NLRP2 and caspase-1 (by Western blot), the expression of NLRP2 mRNA (by real-time polymerase chain reaction) and interleukin-1beta (IL-1β) content (by enzyme-linked immunosorbent assay). Results:The MWT was significantly lower at T 2-4 than at T 0 in group NP, group NP+ siRNA and group NP+ scrRNA ( P<0.05). Compared with group S, the MWT was significantly decreased at T 2-4, the expression of NLRP2 protein and mRNA and caspase-1 was up-regulated, and the content of IL-1β was increased in group NP ( P<0.05). Compared with group NP, the MWT was significantly increased at T 2-4, the expression of NLRP2 protein and mRNA and caspase-1 was down-regulated, and the content IL-1β was decreased in group NP+ siRNA ( P<0.05), and no significant change was found in the parameters mentioned above in group NP+ scrRNA ( P>0.05). Conclusion:NLRP2 in DRG is involved in the development of NP, and the mechanism is related to NLPR2 inflammasomes-induced peripheral neuroinflammation in rats.

MicroRNA-370 Regulates Cellepithelial-Mesenchymal Transition, Migration, Invasion, and Prognosis of Hepatocellular Carcinoma by Targeting GUCD1

PURPOSE: Hepatocellular carcinoma (HCC) is a highly aggressive malignant tumor, the prognosis of which remains poor. Recently, microRNAs have been reported to play crucial functions in multiple tumors, including HCC. However, the molecular mechanisms of miR-370 in HCC still remain largely unknown. The present study focused on the effects of miR-370 on HCC migration, invasion, and epithelial-mesenchymal transition (EMT). MATERIALS AND METHODS: We investigated the key roles and possible regulatory mechanism of miR-370 in regulating HCC metastasis with functional assays, such as transwell assay. Quantitative real-time PCR (qRT-PCR) was used to detect miR-370 and guanylylcyclase domain containing 1 (GUCD1) expression in HCC tissues and cells. Subsequently, we performed transwell assays to determine the functions of miR-370 in HCC cell invasion and migration. Western blot was used to determine protein expressions of relevant genes. Luciferase reporter assays were conducted to confirm the target gene of miR-370. RESULTS: qRT-PCR analysis demonstrated that miR-370 was dramatically downregulated in HCC. Moreover, downregulated miR-370 was found to be associated with poor survival and adverse clinicopathologic characteristics of HCC patients. Transwell assays revealed that miR-370 overexpression dramatically suppressed HCC invasion and migration. Meanwhile, miR-370 restoration prominently inhibited EMT progression in HCC cells. Luciferase reporter assays confirmed GUCD1 as a downstream target gene of miR-370. GUCD1 expression in HCC tissues was prominently increased and inversely correlated with miR-370 expression. Furthermore, GUCD1 was verified as mediating the suppressive influence of miR-370 on cell metastasis and EMT in HCC. CONCLUSION: Taken together, our study confirmed that miR-370 suppressed HCC cell metastasis and EMT via regulating GUCD1. Accordingly, the miR-370/GUCD1 axis may potentially acts as attractive therapeutic targets and novel biomarkers for HCC treatment.

Risk factors for secondary infection in the treatment with anti-neutrophil cytoplasmic antibody-associated vasculitis / 中华肾脏病杂志

Objective To investigate the clinical characteristics and risk factors of secondary infection in patients with anti-neutrophil cytoplasmic antibody-associated vasculitis (AAV).Methods One hundred and eighteen patients newly diagnosed with AAV at the institute of nephrology,Tongji hospital affiliated to Huazhong university of science and technology,from 2012 to 2017,were analyzed retrospectively.Induction therapy included single corticosteroids,combination of corticosteroids with cyclophosphamide and combination of corticosteroids with other immunosuppressive agents.End point was defined as moderate to severe infection which was diagnosed by the clinical and radiological manifestation as well as microbiological evidences.The infection-related survival curve was drawn to reflect the time when the infection occurred.The clinical baseline variables in patients with and without infection were compared.Multivariate Logistic regression model was used to determine the independent predictors of infection.Receiver-operating characteristic curve (ROC) was plotted for evaluating the predictive value of lymphocyte on moderate to severe infection.Results During followup of median 3 months (1-30 months),88 infection episodes were found in 63 (53.4％) patients,of which 54 times (61.4％) occurred within 6 months after treatment,46 times (52.3％) happened within 3 months after treatment.The most common organ of infection was lung (62.5％),and the most common pathogen was bacteria (51.1％).Multivariate Logistic regression model showed that lung involvement (OR=4.44,95％ CI 1.59-12.41),moderate reduction of lymphocyte in follow-up (OR=5.69,95％ CI 2.05-15.85) and severe lymphocyte reduction (OR=36.28,95％CI 3.45-381.17) were independent risk factors of secondary infection in AAV patients (all P ＜ 0.05).ROC curve showed that the area under the curve of lymphocyte as a predictor of severe infection was 0.767 (95％ CI 0.64-0.89,P ＜ 0.05).Based on lymphocyte less than 0.49× 109/L which was the cut-off value for predicting severe infection,the sensitivity and the specificity were 83.9％ and 71.9％,respectively.Conclusions Lung involvement and moderate-severe lymphopenia during follow-up are independent risk factors of secondary infection in AAV patients.Hence,physician should pay more attention to those patients,and adjust treatment in time to avoid the occurrence of infection.

Statins therapy and intracerebral hemorrhage / 国际脑血管病杂志

Despite the extremely high mortality and disability rates of intracerebral hemorrhage,there are currently no effective interventions to improve the outcomes.More and more research shows that statins may be a new method for treating intracerebral hemorrhage.However,there is still much controversy about the relationship between statins therapy and intracerebral hemorrhage.This article reviews the recent studies on the relationship between statins and intracerebral hemorrhage.

Statins therapy and intracerebral hemorrhage / 国际脑血管病杂志

Despite the extremely high mortality and disability rates of intracerebral hemorrhage, there are currently no effective interventions to improve the outcomes. More and more research shows that statins may be a new method for treating intracerebral hemorrhage. However, there is still much controversy about the relationship between statins therapy and intracerebral hemorrhage. This article reviews the recent studies on the relationship between statins and intracerebral hemorrhage.

The effect of microRNA-129 on the migration and invasion in NSCLC cells and its mechanism.

PURPOSE: To investigate the effect of microRNA-129 on the migration and invasion in (Non small cell lung cancer) NSCLC cells and involved mechanism. MATERIALS AND METHODS: A549 cells were cultured and transfected with microRNA-129 (miR-129) mimic. Wound-healing assay and transwell assay were used to analyses the migration and invasion of A549 cells, respectively. The SOX4 expression changes were detected by qRT-PCR and Western blot. Dual-luciferase reporter assay was used to determine the binding site between miR-129 and the 3'-UTR of SOX4. A549 cells, miR-129 up-regulated A549 cells and miR-129 + SOX4 up-regulated A549 cells were treated with TGF-ß in order to induce the epithelial-mesenchymal transition (EMT). The miR-129 and SOX4 after TGF-ß treatment were measured. The EMT markers level changes were detected by western blot. RESULTS: Up-regulation of miR-129 significantly inhibited the migration and invasion of A549 cells. miR-129 mimic transfection could reduce the mRNA and protein level of SOX4. Dual-luciferase reporter assay showed that miR-129 could bind to the 3'-UTR of SOX4. TGF-ß induced EMT accompanied by the reduction of miR-129 and elevating of SOX4. Up-regulation of miR-129 could reverse the EMT marker changes caused by TGF-ß, however, the SOX4 overexpression antagonized this effect of miR-129. CONCLUSIONS: miR-129 could suppress the migration and invasion of A549 cells. The potential mechanism may be that miR-129 inhibit EMT via targeting SOX4.

Effect of hydrogen sulfide on myocardial exogenous apoptotic pathway in a rat model of hemorrhagic shock and resuscitation / 中华麻醉学杂志

Objective To evaluate the effect of hydrogen sulfide on myocardial exogenous apoptotic pathway in a rat model of hemorrhagic shock and resuscitation.Methods Sixty clean-grade healthy male Sprague-Dawley rats,aged 8 weeks,weighing 250-300 g,were divided into 4 groups (n =15 each) using a random number table method:sham operation group (group S),sham operation plus sodium sulphid (NaHS) group (group S+NaHS),hemorrhagic shock group (HS group),and hemorrhagic shock plus sodium sulphid group (group HS+NaHS).Rats only underwent arterial and intravenous puncture in group S.Hemorrhagic shock was induced by withdrawing blood from the femoral artery until mean arterial pressure (MAP) was reduced to 35-40 mmHg within 10 min and maintained for 1.5 h.NaHS 28 μmol/kg was intraperitoneally injected at 10 min before resuscitation in group HS+NaHS.The equal volume of NaHS was administered at the same time in group S+NaHS.Immediately before blood letting and at 0,1.5,2,3,4 and 6 h after blood letting (T1-5),MAP was recorded and blood samples were collected from the femoral vein for determination of serum creatine kinase (CK) and lactate dehydrogenase (LDH) concentrations by chemical colorimetry.Rats were then sacrificed and hearts were removed for examination of the pathological changes of myocardial tissues (with a light microscope) and for determination of cell apoptosis (by TUNEL),expression of caspase-3 and caspase-8 (by Western blot) and expression of Fas and FasL (by immunohistochemistry).Apoptosis index was calculated.Results Compared with group S,MAP was significantly decreased at T1-5,the serum CK and LDH concentrations at T1-5 and apoptosis index at T5 were increased,and the expression of Fas,FasL,caspase-3 and caspase-8 was up-regulated in group HS (P＜ 0.05).Compared with group HS,MAP was significantly increased at T1-3,the serum CK and LDH concentrations at T3-5 and apoptosis index at T5 were decreased,and the expression of Fas,FasL,caspase-3 and caspase-8 was down-regulated in group HS+NaHS (P＜0.05).The pathological changes of myocardial tissues were significantly attenuated in group HS+NaHS when compared with group HS.Concclusion The mechanism by which hydrogen sulfide attenuates myocardial injury induced by hemorrhagic shock and resuscitation is associated with inhibiting the exogenous apoptotic pathway in rats.

Impact of gastric tube diameter on quality of life of esophagus cancer patient after Ivor-Lewis esophagectomy / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the impact of the gastric tube diameter on quality of life of esophagus cancer patients after Ivor-Lewis esophagectomy.</p><p><b>METHODS</b>Clinical and follow-up data of 188 esophageal cancer patients who underwent Ivor-Lewis esophagectomy at Department of Cardio-Thoracic Surgery, Drum Tower Clinical Medicine College, Nanjing Medical University from January 2015 to June 2016 were retrospectively analyzed. Inclusion criteria included age <75 years old, good foundation health situation, no distant metastasis, complete follow-up data for one-year after surgery, and middle-lower esophageal squamous cell carcinoma (ESCC). According to the diameter of gastric tube formed during operation, 92 patients were assigned to narrow gastric tube group (NGT group, ≥2 cm to <4 cm), which were further divided into narrower group (≥2 cm to <3 cm, n=44) and medium narrow group (≥3 cm to <4 cm, n=48); 96 patients were assigned to wide gastric tube group(WGT group, ≥4 cm), which were further divided into medium wide group(≥4 cm to <5 cm, n=50) and wider group(≥5 cm, n=46). Postoperative patients were followed up by telephone or outpatient service for one year and then re-hospitalized to receive associated examinations, including lung function test, esophageal pressure measurement, 24-hour esophageal dynamic pH monitoring (total number of pH<4, number of pH<4 lasting more than 5 minutes, maximum duration of pH<4 and time percentage of pH<4) and dilatation measurement of gastric tube (the diameter measured by CT minus the diameter measured in surgery). During follow-up, postoperative quality of life(QoL) was assessed by questionnaire. These contents were compared and plotted as a chart.</p><p><b>RESULTS</b>There were no statistically significant differences between NGT group and WGT group regard to preoperative baseline information, postoperative pathology and postoperative complications (residual gastric leakage, anastomotic leakage, anastomotic stenosis, pulmonary complications, atrial fibrillation and chylothorax) (all P>0.05). Compared with WGT group, the NGT group had better postoperative lung function, including percentage of vital capacity [(76.4±6.8)% vs. (73.2±7.7)%, t=2.168, P=0.033], percentage of maximal voluntary ventilation [(72.7±6.4)% vs. (69.3±6.8)%, t=2.409, P=0.018] and percentage of forced expiratory volume in the first second [(69.2±5.0)% vs. (66.7±6.2)%, t=2.033, P=0.045], higher plane pressure of anastomotic stoma [(5.4±3.1) mmHg vs. (4.2±2.4) mmHg, t=2.083, P=0.038], greater dilatation of gastric tube [(1.0±0.4) cm vs. (0.5±0.3) cm, t=5.888, P=0.000], milder gastroesophageal reflux according to the indices of 24-hour esophageal dynamic pH monitoring, including the total number of pH<4 (228.3±65.3 vs. 280.8±103.9, t=-2.920,P=0.004), the number of pH<4 lasting more than 5 minutes (19.9±8.5 vs. 30.6±15.6, t=-4.127,P=0.000), the maximum duration of pH<4[(32.5±9.4) minutes vs. (37.9±13.6) minutes, t=-2.232,P=0.028] and the time percentage of pH<4 [(23.4±10.2)% vs. (28.4±10.6)%, t=-2.303, P=0.024]. However, no significant difference was found in the scores of postoperative QoL between the two groups(P=0.051). According to the pairwise comparisons among the four subgroups, narrower group showed better performance on postoperative lung function, plane pressure of anastomotic stoma, the dilatation of gastric tube, indices of 24-hour esophageal dynamic pH monitoring and scores of postoperative QoL as compared to wider group (all P<0.05). There were no statistically significant differences among medium narrow group, medium wide group and wider group. Line charts showed that the larger of the gastric tube diameter, the worse of the postoperative lung function, the more severe of gastroesophageal reflux and the smaller degree of gastric tube dilatation.</p><p><b>CONCLUSION</b>Narrow gastric tube with a diameter of 2-4 cm can improve the postoperative QoL of esophagus cancer patients after Ivor-Lewis esophagectomy without increasing the risk of postoperative complications.</p>

CT-guided percutaneous irreversible electroporation of locally advanced pancreatic cancer: a complications analysis / 中华放射学杂志

Objective To investigate the CT-guided percutaneous irreversible electroporation (IRE) in treating locally advanced pancreatic cancer (LAPC) and providing guidance for its prevention and treatment. Methods We retrospectively analyzed the clinical and imaging data of 17 patients (17 lesions) of LAPC treated with CT-guided IRE in our hospital from July 2015 to June 2016. Complications were documented and reviewed at day 7,30 and 90 follow up as well as during the procedure. The Clavien?Dindo was used for classification. The reasons that induced complications were summarized and to further discuss the prevention and treatment approaches. Results Of 17 patients, 2 patients suffered a transient tachycardia during the procedure. Eleven patients (65%) showed complications at day 7, of which gradeⅠcomplications occurred in 6 cases, including abdominal pain, nausea, vomiting, or a few of inflammatory exudations around the pancreas; four patients have grade Ⅱ complications, along with portal vein thrombosis;one patient showed grade Ⅲ a complications for retroperitoneal infection. With 30 days follow up, the grade Ⅰ complications disappeared, gradeⅡcomplications have not getting better, while grade Ⅲ a complications have been improved. With 90 days follow up,patients with grade Ⅲ a complications getting better; two patients with grade Ⅱ complications didn't show any changes;2 cases progressed to grade Ⅴ, and died of digestive tract bleeding at 82 days and 98 days after procedure. Conclusion CT-guided irreversible electroporation for treating LAPC is a safe ablation approach. Strict patient selection before procedure and make a reasonable prevention and treatment measures can reduce the complications.