The purpose of the present study was to explore whether substance P (SP) modulates the response mediated by GABAA receptors. Experiments were carried out on cultured hippocampal pyramidal neurons of rats. GABA-activated inward currents were recorded using the whole-cell-patch-clamp techique. The majority of the neurons examined (66/92, 72%) were sensitive to both GABA and SP. When the neurons were treated with SP prior to application of GABA, the GABA-activated current (IGABA) was inhibited markedly, which was concentration-dependent and could be blocked by spantide, an NK1 receptor antagonist. With 10－8, 10－7, 10－6 and 10－5 mol/L SP, IGABA was inhibited by 18%, 24.8%, 25.9% and 28% respectively. Intracellular application of H7, a potent inhibitor of PKC, abolished inhibition of IGABA by SP, suggesting that the inhibition of IGABA by SP may be a result of intracellular phosphorylation of the GABAA receptor.

Objective To evaluate the efficacy of argon surper-cryosurgery system (ASSS) in the treatment of huge hepatic multi-nidus carcinoma at advanced stage in cases of which conventional surgery is impossible. Methods Twenty-two patients with huge hepatic multi-nidus carcinoma at advanced stage, incurable with conventional surgery, were treated by ASSS in combination with entire or reduced-size excision. In terms of combined treatment, 1 patient received reduced-size operation and portal vein chemotherapeutic pump, 2 with entire excision and portal vein chemotherapeutic pump, 1 with entire excision and liver artery chemotherapeutic pump, 3 with only liver artery chemotherapeutic pump, and 6 with portal vein chemotherapeutic pump. Nine patients were simply treated by ASSS. Results The surgical procedures went smoothly in all cases and the complications, such as bleeding at the puncture aperturae, thorax effusion, bile leakage and liver cataphora, were successfully managed. The follow-up study was performed and all patients surived with the longest survival of 11 months up till now. Conclusions ASSS offers a new treatment method for huge hepatic multi-nidus carcinoma at advanced stage, but the indications must be carefully observed, and the surgical skills and prevention of complication are of great importance.

Objective To evaluate the efficacy of argon surper-cryosurgery system (ASSS) in the treatment of huge hepatic multi-nidus carcinoma at advanced stage in cases of which conventional surgery is impossible. Methods Twenty-two patients with huge hepatic multi-nidus carcinoma at advanced stage, incurable with conventional surgery, were treated by ASSS in combination with entire or reduced-size excision. In terms of combined treatment, 1 patient received reduced-size operation and portal vein chemotherapeutic pump, 2 with entire excision and portal vein chemotherapeutic pump, 1 with entire excision and liver artery chemotherapeutic pump, 3 with only liver artery chemotherapeutic pump, and 6 with portal vein chemotherapeutic pump. Nine patients were simply treated by ASSS. Results The surgical procedures went smoothly in all cases and the complications, such as bleeding at the puncture aperturae, thorax effusion, bile leakage and liver cataphora, were successfully managed. The follow-up study was performed and all patients surived with the longest survival of 11 months up till now. Conclusions ASSS offers a new treatment method for huge hepatic multi-nidus carcinoma at advanced stage, but the indications must be carefully observed, and the surgical skills and prevention of complication are of great importance.

OBJECTIVETo observe effects of electroacupuncture (EA) on gastric blood perfusion and search for a study method reflecting changes of blood perfusion.

METHODSOn the basis of establishing a new gastric ischemia-reperfusion model which was made by ligature of gastric right artery, EA was given at "Zusanli" (ST 36) and the gastric blood perfusion images were displayed by a laser Doppler blood perfusion imager (LDPI), and effects of EA on gastric blood perfusion were analyzed.

RESULTS(1) The image of the gastric ischemia-reperfusion is clear. The ischemic degree of the stomach in the EA group was lower than the control group within 30 min of ischemia, and recovery of blood flow after perfusion in the EA group was better than the control group; (2) at 25 min of ligature, there was a significant difference in changes of blood perfusion between the EA group [(-0.50 +/- 0.18) PU] and the control group [(-0.90 +/- 0.16) PU], P < 0.05.

CONCLUSIONEA can improve and promote gastric blood supply in the gastric ischemia-reperfusion model, which can be used for acupuncture and moxibustion experimental study.

Animals ; Electroacupuncture ; Female ; Laser-Doppler Flowmetry ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; therapy ; Stomach ; blood supply

OBJECTIVETo evaluate the impact of specific siRNA on survivin gene in transfected leukemia cells.

METHODThe small interfering RNA (siRNA) targeted survivin mRNA was synthesized in vitro and was transfected into K562 cell by Hiperfect into human leukemia cell line K562, which has high survivin expression level. The level of survivin mRNA expression was determined by quantitative reverse transcription polymerase chain reaction (RT-PCR) with SYBR GREEN I. The apoptosis index of cytotrophoblasts were determined and analyzed by FCM (Annexin V-FITC/PI staining methods). The cell proliferation was examined by MTT at 48 h and 72 h after transfection.

RESULTThe level of mRNA expression was significantly inhibited by the siRNA 48 h and 72 h after transfection, the suppression rate of survivin mRNA separately reached 85.21%, 94.35% mensurated by quantitative RT-PCR with SYBR GREEN I, cell proliferation was inhibited significantly by 45.02% and 50.88%, respectively, the apoptotic rate detected by Annexin V-FITC assay reached 12.28%and 21.55%, respectively.

CONCLUSIONThe chemosynthesized siRNA targeting survivin could significantly down-regulate survivin mRNA. Survivin siRNA was able to inhibit the proliferation of leukemia cell line K562. Survivin may become a new target for leukemia gene therapy.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Gene Silencing ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; K562 Cells ; RNA, Small Interfering ; pharmacology ; Transfection

OBJECTIVETo explore the effect of ITF2357, a novel histone deacetylase (HDAC) inhibitor, on the growth, differentiation and apoptosis of acute myeloid leukemic (AML) cells and its mechanism.

METHODSAML cell lines kasumi-1 cells as a model for AML1-ETO positive, and THP1 cells for AML1-ETO negative, the leukemic cells proliferation was analyzed by MTT assay, expression of myeloid-specific differentiation antigen and cell cycle by flow cytometry, cell apoptosis by annexin V staining and flow cytometry. AML1-ETO, acetyl-histone, and caspase protein was analyzed by Western blot.

RESULTS0.5 µmol/L ITF2357 treatment significantly inhibited kasumi-1 cells proliferation, with the 48 h half inhibitory concentration (IC(50)) of 0.1 µmol/L. The initial inhibitory concentration of THP1 cell line was 5 µmol/L. ITF 2357 induced apoptosis of kasumi-1 cells in a time- and dose-dependent manner. A dose-dependent increase in early apoptosis occurred at 24 hours treatment and in late apoptosis at 48 hours treatment by ITF2357. Early apoptosis cells increased from (1.44 ± 1.52)% to (24.51 ± 5.79)%. Late apoptosis cells increased from (2.37 ± 2.8)% to (63.66 ± 1.56)%. ITF2357 induced AML1-ETO degradation by caspase-dependent pathway. 0.25 µmol/L ITF2357 induced a time- and dose-dependent increase in expression of myeloid cell surface protein CD13 and CD15. 5 µmol/L ITF2357 blocked the cells at G(0)/G(1) phase, G(0)/G(1) cells increased from (39.69 ± 6.56)% to (79.2 ± 6.51)% and s-phase cells declined from (60.12 ± 3.29)% to (18.97 ± 6.62)%. Kasumi-1 cells incubated with 0.5 µmol/L of ITF2357, AML1-ETO protein began to decrease at 24 hours and could hardly be detected at 96 hours. ITF2357 induced AML1/ETO degradation through a caspase-dependent mechanism. At the same time, acetylated H3 and H4 increased.

CONCLUSIONLow-dose HDAC inhibitor ITF2357 can effectively inhibit the AML cells proliferation, especially for AML1-ETO positive AML cells. It inhibits Kasumi-1 cells proliferation degradation of AML1-ETO protein expression, blocks the cells at G(0)/G(1) phase, and induces apoptosis and differentiation of the cells.

Acetylation ; Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Histone Deacetylase Inhibitors ; pharmacology ; Histones ; metabolism ; Humans ; Hydroxamic Acids ; pharmacology ; Oncogene Proteins, Fusion ; metabolism

OBJECTIVETo investigate the expression of Oct4, Sox2, Nanog, SMO, beta-Catenin and Wnt5b mRNA in four hepatocellular carcinoma cell lines of SMMC-7721, Bel-7402, HepG2, MHCC-97 and normal hepatocellular cell line of L02, and to compare the response of these cell lines to all-trans retinoic acid.

METHODSRT-PCR was used to detect expression of Oct4, Sox2, Nanog, SMO, beta-Catenin and Wnt5b mRNA in four hepatocellular carcinoma cell lines and normal hepatocellular cell line. Real time-PCR was used to quantify the expression of the genes.

RESULTSThere are different levels of expression of the stem cell-related gene in hepatocellular carcinoma cell lines and control cell line (P less than 0.05). There are significant differences in HepG2 and L-02 for the response to all-trans retinoic acid (P less than 0.05).

CONCLUSIONSThe stem cell-related genes are differentially expressed in different hepatocellular carcinoma cell lines.

Carcinoma, Hepatocellular ; metabolism ; pathology ; DNA Methylation ; Gene Expression Regulation, Neoplastic ; Hep G2 Cells ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Nanog Homeobox Protein ; Octamer Transcription Factor-3 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; SOXB1 Transcription Factors ; genetics ; metabolism ; Signal Transduction ; Smoothened Receptor ; Stem Cells ; drug effects ; metabolism ; Tretinoin ; pharmacology ; Wnt Proteins ; genetics ; metabolism ; beta Catenin ; genetics ; metabolism

Objective To study whether the presence of gastric pepsin in the sputum might be used as a reliable criteria of laryngopharyngeal reflux. Methods Fifty-six patients with the symptoms of laryngopharyngitis and fifteen healthy people were recruited. Fifty-six patients were divided into laryngopharyngeal reflux group and chronic laryngitis group by the reflux symptom index (RSI), by the reflux finding score (RFS) and by their treating experiment taking omeprazole 20 mg bid for 2 weeks. Sputum in all three groups was obtained in the morning. Pepsin in the sputum was measured by enzyme-linked immunoadsorbent assay. Results The positive rate of pepsin in sputum among LPR group, chronic laryngopharyngitis group and normal group were 93.8% (30/32), 75.0% (18/24), 20.0% (3/15) respectively, and the median concentration of pepsin were 5. 3 [1.3;53.4] ng/ml, 0.8 [0.1;17.2] ng/ml, 0.0 [0.0;0.0] ng/ml (H=23.29, P=0.000). Compared with co-diagnosis as gold standard, the sensitivity of RSI, RFS treating experiment and the pepsin immunoassay was 59.4%, 84.4%, 81.3% and 93.8%, and the specificity of those was 87.2%, 61.5%, 95. 8% and 46.2% respectively. Conclusions Detection of pepsin in sputum by immunoassay might provide a high sensitive, noninvasive method for laryngopharyngeal reflux.

OBJECTIVETo investigate and summarize the experience in clinical presentation, diagnosis and treatment of portal vein thrombosis after orthotopic liver transplantation (OLT).

METHODSThe clinical data of 402 patients who underwent OLT from January 2003 to February 2007 were reviewed. A retrospective study was performed on etiology, prognosis and treatment in 9 cases of portal vein thrombosis after OLT.

RESULTSAll of the 9 cases received anticoagulant and antiaggregation therapy, within whom one underwent percutaneous transluminal angioplasty and stent placement, one underwent retransplantation after failure of thrombolysis therapy, and one received surgical embolectomy. Six patients died of multiple organ failure on 9th, 30th, 34th, 40th, 48th, 6 2nd days, respectively, while 3 patients survived.

CONCLUSIONSThe major risk factors of portal vein thrombosis after OLT were pathological changes in portal vein, abnormal blood stream dynamics, hypercoagulable status and improper surgical technique. Prophylactic intervention to patients with high risk factors, early diagnosis and aggressive comprehensive therapy on portal vein thrombosis patients are essential to improve prognosis.

Adult ; Female ; Humans ; Liver Transplantation ; adverse effects ; Male ; Middle Aged ; Portal Vein ; Postoperative Complications ; diagnosis ; etiology ; therapy ; Prognosis ; Retrospective Studies ; Venous Thrombosis ; diagnosis ; etiology ; therapy

Objective To investigate the clinical data, risk factors for bleeding following endoscopic treatment of colorectal polyps (diameter ≥ 1.0 cm) and feature analysis for carcinogenesis of adenomatous polyps. Method It was analyzed retrospectively that the clinical data, endoscopic characteristics, risk factors for bleeding and features for carcinogenesis of adenomatous polyps in 741 patients with a total of 884 colorectal polyps (diameter ≥ 1.0 cm) of our hospital from January 1, 2014 to January 1, 2016, which were resected under endoscope. Result Univariate analysis of colorectal polyps resected under endoscope on intraoperative and delayed bleeding, we found that gender (P = 0.017), location (P = 0.011), size (P = 0.004), lobulated or not (P = 0.010), resection methods under endoscope (P = 0.029) were statistically significant deviation between the two groups; taking the polyp as observation unit,multivariate Logistic regression analysis of their clinical data and endoscopic characteristics, we found gender (P = 0.012, OR^ = 2.671, 95% CI = 1.246 ~ 5.728) was an independent risk factor, men are more prone to bleeding than women; it was a protective factor for the location of polyps is sigmoid colon compared to rectum (P = 0.011, OR^ = 0.348, 95% CI = 0.154 ~ 0.786), and it was a risk factor for the polyps whose diameters ≥ 3.0 cm compared to among 1.0 ~ 1.9 cm. Univariate analysis of the features for carcinogenesis of colorectal adenomatous polyps, we found that surface lobulated or not (P = 0.001), surface smooth or not (P = 0.017), Yamada classification (P = 0.008) were statistically significant deviation, and the logistic regression analysis identified surface lobulation (P = 0.001, OR^ = 6.556, 95% CI = 2.326 ~ 18.475) was an independent risk factor for carcinogenesis of adenomatous polyps. Conclusion Resection of colorectal polyps under endoscope is a safe treatment method. Taking the polyp as observation unit, gender is an independent risk factor of colorectal polyps resected under endoscope on intraoperative and delayed bleeding, men are more prone to bleeding than women; the larger the diameters of polyps are, the higher the bleeding rates could be. Surface lobulation of colorectal adenomatous polyps are more likely lead to carcinogenesis.