Objective:To investigate whether hydrogen sulfide (H 2S) protects against intestinal ischemia/reperfusion (I/R) injury in rats by regulating c-Jun N-terminal kinase/activator protein-1 (JNK/AP-1) signaling pathway. Methods:Thirty male Wistar rats were divided into sham operated group (Sham group), I/R group, and H 2S donor sodium hydrosulfide (NaHS) intervention group (I/R+NaHS group), with 10 rats in each group. The I/R injury model was established by blocking the superior mesenteric artery with a non-traumatic vascular clip, with 60 minutes of ischemia followed by 120 minutes of reperfusion. In the I/R+NaHS group, 100 μmol/kg of NaHS was injected through the tail vein 10 minutes before reperfusion, followed by continuous infusion of 1.07 mmol·kg -1·h -1 until the end of the 120-minute reperfusion period. Plasma H 2S concentration was measured using a sensitive sulfur electrode. Malondialdehyde (MDA) and superoxide dismutase (SOD) levels in the small intestine tissue were assayed spectrophotometrically. Histological sections of the small intestine were stained with hematoxylin-eosin (HE) staining and scored using the Chiu scoring system to assess the degree of intestinal mucosal injury. Western blotting was used to detect the protein expressions of phosphated-JNK (p-JNK), JNK, AP-1, and BCL-2 in the small intestine tissue. Results:Compared with the Sham group, the I/R group exhibited damage to the lamina propria, hemorrhage, and ulceration, with a significantly higher Chiu score (4.80±0.63 vs. 0.70±0.09, P < 0.01); plasma H 2S concentration and SOD activity in the ileum tissue were significantly reduced [H 2S (μmol/L): 17.29±1.40 vs. 34.62±1.48, SOD (kU/g): 5.38±0.93 vs. 20.56±1.85, both P < 0.01], while MDA level was significantly elevated (μmol/g: 16.06±1.71 vs. 4.80±0.92, P < 0.01); expression of BCL-2 protein in the ileum tissue was significantly down-regulated (BCL-2/β-actin: 0.32±0.06 vs. 0.79±0.05, P < 0.01), while expressions of p-JNK and AP-1 proteins were significantly up-regulated (p-JNK/β-actin: 0.69±0.03 vs. 0.10±0.03, AP-1/β-actin: 0.82±0.02 vs. 0.22±0.02, both P < 0.01). Compared with the I/R group, the I/R+NaHS group showed moderate separation between the epithelial and lamina propria layers, with partial damage to the tips of the villi; the Chiu score was significantly lower (2.90±0.56 vs. 4.80±0.63, P < 0.01); plasma H 2S concentration and SOD activity in the ileum tissue were significantly increased [H 2S (μmol/L): 24.48±1.84 vs. 17.29±1.40, SOD (kU/g): 10.29±1.26 vs. 5.38±0.93, both P < 0.01], while MDA level was significantly reduced (μmol/g: 7.88±1.01 vs. 16.06±1.71, P < 0.01); expression of BCL-2 protein in the ileum tissue was significantly up-regulated (BCL-2/β-actin: 0.44±0.06 vs. 0.32±0.06, P < 0.01), while expressions of p-JNK and AP-1 proteins were significantly down-regulated (p-JNK/β-actin: 0.54±0.05 vs. 0.69±0.03, AP-1/β-actin: 0.66±0.04 vs. 0.82±0.02, both P < 0.01). There was no statistically significant difference in the expression of JNK in the ileum tissue among the Sham group, I/R group, and I/R+NaHS group (JNK/β-actin: 0.63±0.02, 0.66±0.02, 0.64±0.02, respectively, P > 0.05). Conclusion:H 2S exerts a protective effect on intestinal I/R injury in rats by down-regulate the expression of the JNK/AP-1 signaling pathway, as well as reducing oxidative stress levels.

Objective:To explore the effect of hydrogen sulfide (H 2S) on expression of phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) signal pathway in rats with intestinal ischemia/reperfusion (IRI) injury. Methods:Thirty male Wistar rats were divided into sham operation group (Sham group), IRI group, and H 2S precursor sodium hydrosuphide (NaHS) intervention group (IRI+NaHS group) by random number table method, with 10 rats in each group. The animal model of IRI was established by 60 minutes superior mesenteric artery (SMA) blockage with non-invasive vascular clamp and 120 minutes reflow. SMA was dissociated and peritoneum cavity was closed in Sham group. The rats in IRI+NaHS group was received NaHS (100 μmol/kg bolus+1.07 mmol·kg -1·h -1 infusion) 10 minutes prior to the onset of reperfusion, while the rats in IRI group and Sham group were received equal volume of normal sodium. Blood in vena cava was collected. H 2S was detected by sensitive sulfide electrode. Rats were sacrificed after blood collection. Histopathology change was observed by hematoxylin-eosin (HE) staining, ileal pathological score was studied by Chiu score. The protein expressions of phosphated Akt (p-Akt), Akt, PI3K, cleaved caspase-9, mammalian target of rapamycin (mTOR) were determined by Western Blot. Results:Compared with the Sham group, there was intestinal mucosa structure disorder edema and shedding villous fracture in the IRI group. Ileal pathological score in IRI group was significantly increased (4.21±0.15 vs. 0.15±0.03, P < 0.01), while plasma H 2S in IRI group was significantly decreased (μmol/L: 26.72±3.17 vs. 38.34±5.24, P < 0.01). Ileal p-Akt, PI3K, caspase-9 and mTOR protein in IRI group were significantly increased (p-Akt/GAPDH: 2.67±0.12 vs. 0.24±0.05, PI3K/GAPDH: 1.42±0.07 vs. 0.57±0.08, caspase-9/GAPDH: 4.23±0.61 vs. 0.13±0.02, mTOR/GAPDH: 2.17±0.23 vs. 0.23±0.02, all P < 0.01). Compared with the IRI group, pathological changes of intestinal mucosa in the IRI+NaHS group was improved, ileal pathological score was significantly decreased (1.56±0.02 vs. 4.21±0.15, P < 0.01), plasma H 2S was significantly increased (μmol/L: 32.36±2.45 vs. 26.72±3.17, P < 0.01) and ileal p-Akt, PI3K were significantly increased (p-Akt/GAPDH: 5.12±0.08 vs. 2.67±0.12, PI3K/GAPDH: 3.14±0.05 vs. 1.42±0.07, both P < 0.01), while caspase-9, mTOR in IRI+NaHS group were significantly decreased (caspase-9/GAPDH: 2.12±0.24 vs. 4.23±0.61, mTOR/GAPDH: 1.37±0.28 vs. 2.17±0.23, both P < 0.01). Conclusion:H 2S attenuates intestinal injury in IRI rats by up-regulating PI3K/Akt signal pathway and down-regulating caspase-9 and mTOR.

Objective: To investigate the effect of three-dimensional conformal radiotherapy (3D-CRT) combined with PC chemotherapy (paclitaxel + carboplatin) on non-small cell lung cancer (NSCLC) patients and the serum levels of CA125, tissue inhibitor of metalloproteinase-1 (TIMP-1), serum amyloid A (SAA) and T-lymphocyte subsets. Methods: A total of 100 patients with NSCLC treated in Affiliated Hospital of Guangdong Medical University from May 2015 to December 2017 were selected as the study subjects. They were divided into control group and observation group according to random number table method, with 50 cases in each group. The observation group was treated with 3D-CRT combined with PC chemotherapy, while the control group was treated with PC chemotherapy. The two groups were treated for 4 cycles. The therapeutic effect, serum CA125, TIMP-1, SAA, T-lymphocyte subsets and adverse reactions were compared between the two groups. Results: Four cases were lost to follow-up both in the two groups. The overall response rate in the observation group (43.48%, 20/46) was higher than that in the control group (23.91%, 11/46; χ²=3.941, P=0.047). The serum levels of CA125, TIMP-1 and SAA of the two groups had no significant difference before treatment, and the levels of these indexes decreased after treatment. The serum levels of CA125, TIMP-1 and SAA in the observation group after treatment were (12.31±1.13) U/ml, (275.31±13.69) pg/ml and (47.21±7.21) mg/L, which were lower than those in the control group [(30.36±1.98) U/ml, (320.36±17.23) pg/ml, (65.92±8.36) mg/L], with significant differences (t=53.699, P<0.001; t=13.884, P<0.001; t=11.495, P<0.001). The levels of CD3⁺ , CD4⁺ , CD8⁺ and CD4⁺ /CD8⁺ of the two groups had no significant difference before treatment, and the levels of these indexes decreased after treatment. The levels of CD3⁺ , CD4⁺ , CD8⁺ and CD4⁺ /CD8⁺ in the observation group were (35.27±10.31)%, (20.27±6.72)%, (15.89±3.37)% and 0.91±0.37, which were higher than those in the control group [(30.77±9.27)%, (15.27±5.73)%, (12.02±2.69)% and 0.75±0.39], with significant differences (t=2.201, P=0.030; t=3.840, P<0.001; t=6.087, P<0.001; t=2.019, P=0.047). There were no significant differences in the adverse reactions such as nausea and vomiting [63.04% (29/46) vs. 43.48% (20/46); χ²=3.537, P=0.060], phlebitis [6.52% (3/46) vs. 4.35% (2/46); χ²=0.000, P>0.999], abnormal liver function [6.52% (3/46) vs. 2.17% (1/46); χ²=0.261, P=0.609] and myelosuppression [8.70% (4/46) vs. 6.52% (3/46); χ²=0.000, P>0.999] between the observation group and the control group. Conclusion For patients with NSCLC, 3D-CRT combined with PC chemotherapy can improve the overall response rate, decrease the levels of serum CA125, TIMP-1 and SAA, and improve the immune function of patients. The therapeutic effect is remarkable and the safety is good. The therapeutic scheme is suitable for the treatment of NSCLC.

Objective To retrospectively analyze and compare the clinical application value of core-needle biopsy (CNB) histology and fine needle aspiration (FNA) cytology in diagnosing malignant thyroid nodules. Methods A total of 134 patients with 137 thyroid nodules (93 malignant nodules and 44 benign nodules) were included in this study. Under ultrasound guidance, successive use of 22 G fine needle and18 G core-needle to puncture each nodule was performed for sampling of thyroid nodule. Surgical findings and pathological manifestations were compared with clinical follow-up results. The success rate of sampling and the diagnostic accuracy, sensitivity as well as specificity for malignant thyroid nodules were compared among FNA, CNB, and CNB/FNA. Results The success rate of puncture sampling with FNA, CNB and FNA/CNB for thyroid nodules was 89.1％, 97.8％ and 100％ respectively. For malignant thyroid nodules, the diagnostic accuracy of FNA, CNB and FNA/CNB was 79.6％, 91.9％ and 96.4％ respectively, the sensitivity was 81.7％, 94.6％ and 97.8％ respectively, and the specificity was 75.0％, 86.4％ and 93.2％ respectively. The success rate of puncture sampling by using CNB or FNA/CNB was significantly higher than that by using FNA (P<0.01), moreover, the diagnostic accuracy and sensitivity for malignant thyroid nodules by using CNB or FNA/CNB was also remarkably higher than those by using FNA (P<0.01) . Conclusion In making diagnosis of malignant thyroid nodules, CNB is accurate, safe and reliable. CNB can be used as a complementary or alternative technique to FNA in clinical practice.

OBJECTIVE:To observe the clinical efficacy of sitagliptin combined with benazepril in the treatment of diabetic nephropathy(DN). METHODS:Sixty DN patients admitted to our hospital during Sept. 2014-Jun. 2015 were divided into sitagliptin group,benazepril group,drug combination group according to random number table,with 20 cases in each group. Based on routine treatment,sitagliptin group was given sitagliptin 100 mg orally,qd;benazepril group was given Benazepril 10 mg orally,qd;drug combination group was given sitagliptin 100 mg+benazepril 10 mg orally,qd. The drug dosage would be doubled if the blood pressure of patients in 3 groups had not yet reached the standard. Treatment course of 3 groups lasted for 12 weeks. The levels of 24 h urine protein,IL-6 and Cys-C were measured in 3 groups before and after treatment. Clinical efficacies and the occurrence of ADR were observed. RESULTS:Total response rate of drug combination group(90.00%)was significantly higher than those of sitagliptin group (65.00%)and benazepril group(70.00%);there was statistically significance(P<0.05). After treatment,the levels of 24 h urine protein,IL-6 and Cys-C in 3 groups were significantly lowered,compared to before treatment;those of drug combination group was significantly lower than those of other 2 groups;there was statistically significance(P<0.05). There was no statistical significance in above indexes between sitagliptin group and benazepril group(P>0.05). No obvious ADR was found in 3 groups during treatment. CONCLUSIONS:Both sitagliptin and benazepril can decrease the levels of 24 h urine protein,IL-6 and Cys-C,while drug combination shows better effect and clinical response rate,and does not influence the safety of drug use.

Objective To observe the correlation between CD4+ CD29+ T cells and metastasis and radiotherapy for patients with pulmonary adenocarcinoma. Method Seventy-one patients with lung adenocarcinoma, 93 patients with lung adenocarcinoma ,76 cases of chronic obstructive pulmonary disease (COPD),63 cases of healthy volunteers were enrolled. Frequencies of blood CD4+ CD29+ T cells and their intracellular necrosis factor alpha(TNF-α)and interleukin 1(IL-1)were compared. Compare TNF-α,IL-1,integrin beta 1 and vascular endothelial growth factor(VEGF)levels in the patients with transferred pulmonary adenocarcinoma or with non-transferred pulmonary adenocarcinoma and their changes with the treatment of radiotherapy. Results the patients with lung adenocarcinoma and non lung adenocarcinoma were significantly higher than that of COPD and health group,and patients with lung adenocarcinoma is significantly higher than patients with non lung adenocarcinoma (P<0.05);Integrin beta 1,VEGF and CD4+CD29+T cells,TNF-αand IL-1 level in patients with lung adeno-carcinoma metastasis were significantly higher than non-transferred group(P < 0.05);After radiotherapy,CD4+CD29+T cells,TNF-αand IL-1 in patients with lung adenocarcinoma were significantly lower than before(P<0.05);CD4+ CD29+ T cells,TNF alpha and IL-1 with integrin beta 1 and VEGF had significantly positive correlations. Conclusion CD4+CD29+T cells and cytokines increase significantly in the blood of patients with lung adenocarci-noma,and are related to the prognosis of metastasis and radiation therapy,which has important clinical significance.

Background and objective Lung cancer is the leading cancer-related death worldwide.Patients with lung cancer mainly died of tumor metastasis and invasion.Protein kinase CK2 is an ubiquitous sefine/threonine protein kinase and is frequently upregulated in various human tumors.This study aims to explore the effect and molecular mechanism of the invasion and migration of lung adenocarcinoma A549 cells after knock-down of CK2α expression.Methods The pSilencerTM 4.1-siCK2α-eGFP oflentiviral-me-diated shRNA was constructed.The expression of CK2α was knock-downed,and a stable A549 cell line was established.The invasion and migration ofA549 cell line was detected through Transwell and Boyden chamber assays.The protein expression of the PI3K/Akt signaling pathway and mesenchymal-to-epithelial transition (EMT) was evaluated using Westem blot analysis.Results The invasion and migration of A549 cells were significantly inhibited after the knockdown of CK2α expression compared with that in the control group,p-PTEN,Akt,p-Akt473,p-Akt308,p-PDK1,p-c-Raf,and p-GSK-3β were significantly downregnlated,whereas PTEN was upregulated.Moreover,vimentin,β-catenin,Snail,MMP2,and MMP9 were significantly downregnlated after reducing the CK2α expression.Conclusion CK2α might regulate the invasion and migration of A549 cells through the PI3K/Akt/GSK-3β/Snail signaling pathway,which controls EMT in lung adenocarcinoma.

AIM:To investigate the effect of hydrogen sulfide (H2S) on the expression of MAPKs and ileal epithelial cell apoptosis in the rats with intestinal ischemia-reperfusion ( I/R) injury.METHODS:Healthy female Wistar rats were randomly divided into sham operation group, I/R group, I/R+sodium hydrosulfide ( NaHS) group .The animal model of intestinal ischemia reperfusion was established.Apoptosis index ( AI) of ileal epithelial cell was measured by TUNEL assay.H2 S was detected by sensitive sulfide electrode.The mRNA expression of ERK, JNK and p38MAPK was detected by RT-PCR.The protein levels of phosphorylation( p)-ERK, p-JNK, p-NF-κB P65 and p38MAPK were deter-mined by Western blot.RESULTS:H2 S, ERK mRNA and p-ERK in I/R group were significantly higher than those in I/R+NaHS group and sham group while JNK mRNA, p38MAPK mRNA, p-JNK, p-p38MAPK, p-NF-κB P65 and AI were predominantly higher than those in I/R+NaHS group and sham group (P<0.01).CONCLUSION:H2S attenuates ileal epithelial cell apoptosis in the rats with intestinal I/R injury by down-regulating ERK mRNA p-ERk and up-regulating JNK mRNA, p38MAPK mRNA and phosphorylation of JNK, p38MAPK and NF-κB P65.

Objective To study the change in endogenous hydrogen sulfide (H2S) in patients with acute pancreatitis and its relationship to coagulation function. Methods A prospective case control study was conducted. Forty patients with mild acute pancreatitis (MAP group) and 40 with severe acute pancreatitis (SAP group) admitted to Yiwu Central Hospital in Zhejiang Province from December 2002 to March 2015 were enrolled. Forty healthy persons served as control (healthy control group). Blood was collected to determine the levels of H2S, blood coagulation factor Ⅷ (FⅧ), von Willebrand factor (vWF), plasminogen (PLG), antithrombin (AT), platelet count (PLT), tissue factor (TF), tumor necrosis factor-α (TNF-α), and protease activated receptor-1 (PAR-1). The correlations among the above parameters were analyzed. Results There was no statistical significance in sex, age, body weight and time of disease among three groups, indicating it was comparable among the groups. Compared with healthy control group, the levels of H2S, FⅧ, vWF, TF, TNF-α, and PAR-1 in MAP and SAP groups were significantly elevated [H2S (μmol/L): 67.42±6.34, 112.47±12.69 vs. 42.57±4.18, FⅧ: (67.5±5.8)%, (82.3±4.7)% vs. (57.2±6.4)%, vWF: (112.6±9.7)%, (142.5±12.5)% vs. (76.4±8.2)%, TF (ng/L): 45.27±4.34, 64.76±6.25 vs. 18.15±1.89, TNF-α (ng/L): 197.67±13.62, 324.72±25.54 vs. 20.08±2.57, PAR-1 (fluorescence intensity): 32.16±4.43, 56.12±7.07 vs. 12.27±2.12, all P < 0.01], and PLG and AT activity were significantly decreased [PLG: (52.4±4.7)%, (36.7±3.2)% vs. (62.1±5.6)%, AT: (43.2±6.9)%, (35.5±5.4)% vs. (53.6±6.1)%, all P < 0.01]. The changes in the parameters in SAP group were more remarkable than those in MAP group (all P < 0.01). PLT in SAP group was significantly lower than that in healthy control and MAP groups (×109/L: 8.5±1.1 vs. 15.7±2.8, 12.4±1.9, both P < 0.01). H2S was positively correlated with FⅧ, vWF, TF, TNF-α, and PAR-1 (r value was 0.56, 0.61, 0.72, 0.66, 0.64, respectively, all P < 0.01), and it was negatively correlated with PLG and AT (r value was -0.64, -0.57, both P < 0.01). Conclusion As an inflammatory factor, endogenous H2S deteriorates coagulation function in patients with acute pancreatitis by up-regulating TF, TNF-α, and PAR-1.

OBJECTIVE:To establish the method for the residues determination of 5 organic solvents in teicoplanin raw materi-al and injection. METHODS:Headspace GC was performed on the column with 6% cyanopropylphenyl-94% dimethyl polysiloxane (DB-624)as the stationary phase capillary column,the carrier gas was nitrogen,using the temperature program. The temperature of inlet was 200 ℃,detector was hydrogen flame ionization detector with the flow rate of 1 ml/min,split ratio was 40 ∶ 1 and the vol-ume was 1 ml. RESULTS:Good linearity of ethanol,acetone,ethyl acetate,tetrahydrofuran and triethylamine were obtained(r were 0.999 0-0.999 3);the average recoveries were respectively 95.6%,97.0%,103.2%,94.3%and 98.2%(RSD were 2.1%-4.9%, n=9);RSDs of precision and repeatability tests ≤2.6%;and the minimum detectable concentration were respectively 2,2,2,0.7 and 0.3 μg/ml. CONCLUSIONS:The established method is rapid,sensitive and accurate,and can be used for the residues determi-nation of organic solvents in teicoplanin raw materials and injection.