Effect of strophanthidin (Str) on intracellular calcium concentration ([Ca2+]i) was investigated on isolated ventricular myocytes of guinea pig. Single ventricular myocytes were obtained by enzymatic dissociation technique. Fluorescent signal of [Ca2+]i was detected with confocal microscopy after incubation of cardiomycytes in Tyrode's solution with Fluo3-AM. The result showed that Str increased [Ca2+]i in a concentration-dependent manner. The ventricular myocytes began to round-up into a contracture state once the peak level of [Ca2+]i was achieved in the presence of Str (10 μmol·L-1), but remained no change in the presence of Str (1 and 100 nmol·L-1). Tetrodotoxin (TTX), nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str (1 and 100 nmol·L-1), but had no obvious effects on the action of Str (10 μmol·L-1). The elevation of [Ca2+]i caused by Str at all of the detected concentrations was partially antagonized by rynodine (10 μmol·L-1) or the removal of Ca2+ from Tyrode's solution. In Na+, K+-free Tyrode's solution, the response of cardiomycytes in [Ca2+]i elevation to Str (10 μmol·L-1) was attenuated, while remained no change to Str (1 and 100 nmol·L-1). TTX, nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str at all of the detected concentrations in Na+, K+-free Tyrode's solution. The study suggests that the elevation of [Ca2+]i by Str at the low (nomomolar) concentrations is partially mediated by the extracellular calcium influx through Ca2+ channel or a "slip mode conductance" of TTX sensitive Na+ channel. While the effect of Str at high (micromolar) concentrations was mainly due to the inhibition of Na+, K+-ATPase. Directly triggering the release of intracellular Ca2+ from sarcoplasmic reticulum (SR) by Str may be also involved in the mechanism of [Ca2+]i elevation.

Objective:To investigate the therapeutic effect of carbon dioxide laser tonsillectomy.Method:In this prospective,randomized study, One hundred and two patients were divided into laser group or control group. Patients of laser group were cured with carbon dioxide laser tonsillectom,and the control group was cured with routine method. All operations are executed by one person. Observation index included operation time, hemorrhage in operation, ache after operation, inflammatory reaction of raw surface, repair time of raw surface, rehaemorrhagia and scar.Result:Laser group had advantages of less operation time, less hemorrhage, less ache and less inflammatory reaction of raw surface. Laser group have hemorrhage in operation (7.2±2.1)ml, while control group have hemorrhage in operation (92.0±35.0)ml. Laser group have pseudomembrane early but desquamate late.Conclusion:Carbon dioxide laser tonsillectomy is effective to relieve pain, inflammatory reaction and with less time ,it's an safe , efficient and mini-trauma operation.

Objective To investigate the effect of antisense oligonucleotides (ASODN) targeting livin on the inhibition of livin mRNA and protein expression and the apoptosis of human renal carcinoma cell line 786-O cells. Methods Specific phosphorothioate antisense oligodeoxynucleotides targeting livin were synthesized and then transfected into 786-O cells. The expressions of livin mRNA were detected by RT-PCR. Expression and location of livin protein were observed by confocal laser scanning microscope (CLSM). Apoptosis rate of 786-O cells was investigated by flow cytometer. The activity of Caspase-3 was detected by colorimetric assay. Results After the transfection of ASODN, the expression of livin mRNA was decreased (P

OBJECTIVETo introduce the location and course of S1, S2 sacral nerve root tunnel and to clarify the significance of the anterior aspect of sacral nerve root tunnel on placement of iliosacral screw on the standard lateral sacral view.

METHODSFirstly the data of 2.0 mm slice pelvic axial CT images were imported into Mimics 10.0, and the sacrum, innominate bones, and sacral nerve root tunnels were reconstructed into 3D views respectively, which were rotated to the standard lateral sacral views, pelvic outlet and inlet views. Then the location and course of the S1, S2 sacral nerve root tunnel on each view were observed.

RESULTSThe sacral nerve root tunnel started from the cranial end and anterior aspect of the vertebral canal of the same segment and ended up to the anterior sacral foramen with a direction from cranial-posterior-medial to caudal-anterior-lateral. The tunnel had a lower density than the iliac cortex and greater sciatic notch on the pelvic X-rays,especially on the standard sacral lateral view, on which it showed up as a disrupted are line and required more careful recognition.

CONCLUSIONIt can prevent the iliosacral screw from penetrating the sacral nerve root tunnel and vertebral canal when recognizing the anterior aspect of sacral nerve root tunnel and choosing it as the caudal-posterior boundary of the "safe zone" on the standard lateral sacral view.

Adult ; Aged ; Bone Screws ; Female ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Pelvic Bones ; diagnostic imaging ; injuries ; innervation ; surgery ; Radiography ; Sacrococcygeal Region ; diagnostic imaging ; innervation ; surgery ; Sacrum ; diagnostic imaging ; injuries ; innervation ; surgery ; Spinal Nerve Roots ; diagnostic imaging ; surgery ; Young Adult

Objective To construct replication-deficient recombinant adenoviruses AdHNF4?that co-expresse human hepatocyte nuclear factor 4?(HNF4?) and green fluorescent protein(GFP) gene,and to evaluate the effect of HNF4?up-regulation on hepatocyte gene expression.Methods The HNF4?cDNA was obtained through RT-PCR from human hepatocyte.The recombinant adenoviral plasmid- pAdHNF4?was established using AdEasy system and packed in 293 cells.After transfection of human hepatoma cell lines HepG2 and Hep3B with AdHNF4?,the expression of HNF4?and other liver-associ- ated functional genes were evaluated by RT-PCR and Western blot.Results The recombinant plasmid pAdHNF4?was confirmed by restriction endonuclease digestion and sequencing.GFP expression was observed on the fourth day after packing the linearized pAdHNF4?in 293 cells.Stable transfection of AdHNF4?with a titer of 1?10~(10) efu/ml was obtained after repeated amplification.More than 90% of human hepatoma cells had GFP expression in 72 hours after transfection of AdHNF4?.The expression of HNF4?mRNA and protein were significantly up-regulated compared with the control group(3.4 folds in HepG2 infected with AdHNF4a and 5.2 folds in Hep3B infected with AdHNF4?).Furthermore,the transcriptional expressions of some liver-associated functional genes such as apolipoprotein,cytochrome P450 families,glutamine synthetase,phosphoenolpyruvate carboxykinase and glucose-6-phosphatase also increased after transfection of the virus,and the apoptosis ratio of the cells increased.Conclusions Up- regulating the expression of HNF4?in human hepatoma cells with AdHNF4?could enhance normal liver- specific function.Our study would provide a new idea for the researches on gene regulation of transplan- ted hepatocytes.

Little is known about the non-structural protein 6(NSP6)of rotavirus.This report describes expression of the NSP6 of a group A human rotavirus strain TB-Chen in bacteria,and its immunological properties and cellular distribution.The results showed that the recombinant NSP6(rNSP6)was expressed in high efficiency without any other proteins fused(possesses about 34.2% of total bacterial proteins).rNSP6 elicited mono-specific antibodies in immunized guinea pigs and the antibodies could react with the rNSP6 itself and the viral NSP6 proteins synthesized in SA11-or Wa-infected MA104 cells in Western blot and immunofluorescence assay.The NSP6 distributed evenly in the cytoplasm mainly around the nucleus of virus-infected cells,no viroplasm-like gathering observed;The top amount of NSP6 synthesized in SA11-infected cells or Wa-infected cells could be detected at 12h after infection.This is the first report about the high expression of entire NSP6(without any other proteins fused)in prokaryotic expression system and detection of NSP6 synthesis in virus infected cells by immunofluorescence assay.The results are important to understand the structure,biological properties and further application of the NSP6.

OBJECTIVETo analyze the computed tomography (CT) and magnetic resonance imaging (MRI) findings of small hepatocellular carcinoma to improve the accuracy in the diagnosis.

METHODSThis retrospective analysis involved 41 patients with small hepatocellular carcinoma cases confirmed by pathological examination of the biopsy samples or follow-up. These patients were assessed for CT and MRI findings including lesion size, density or signal intensity, enhancement patterns, and presence of tumor capsules.

RESULTSOn unenhanced CT images, small hepatocellular carcinomas were displayed mainly as low-density masses, and the majority of tumors presented with low signal intensity on T1-weighted unenhanced MR images with increased signal intensity on T2-weighted images in comparison with the surrounding liver parenchyma. Most of tumors showed intense enhancement during the arterial phase (CT in 15 cases and MRI in 13 cases), but some appeared isointense to the liver parenchyma (CT in 4 cases and MRI in 4 cases). In portal and delayed phases, the tumors typically had lower signal intensity than that of the surrounding liver tissues (CT in 25 cases and MRI in 12 cases) with enhancement of the tumor capsules (13 cases).

CONCLUSIONDynamic enhanced scanning can be more informative of the pathology and blood supply of small hepatocellular carcinoma. Early and late arterial phase imaging may help in detecting the small lesions and in making differential diagnosis.

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnosis ; diagnostic imaging ; Diagnosis, Differential ; Female ; Humans ; Image Enhancement ; Liver Neoplasms ; diagnosis ; diagnostic imaging ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed

OBJECTIVETo verify the efficacy on lumbar disc herniation treated with Shu-needle therapy in combination with ozone injection of low concentration.

METHODSOne hundred and thirty cases of lumbar disc herniation were randomized into a Shu-needle therapy group and an acupotomy group, 65 cases in each one. In the Shu-needle therapy group, Shu-needle therapy was used in combination with ozone injection of low concentration. In the acupotomy group, the conventional acupotomy therapy was applied in combination with ozone injection of low concentration. The treatment was given once every 10 days, 3 treatments made one session. After one session treatment, the clinical efficacy of two groups was observed, scores of visual analogue scale (VAS) and Oswestry disability index (ODI) were counted before and after treatment. The long-term efficacy was followed up in half a year.

RESULTSThe clinical curative rate was 69.2% (45/65) and the total effective rate was 96.9% (63/65) in the Shu-needle therapy group. The curative rate was 43.1% (28/65) and the total effective rate was 84.6% (55/65) in the acupotomy group. In comparison, the efficacy of the Shu-needle therapy group was superior to that of the acupotomy group (P < 0.01, P < 0.05). The scores of VAS and ODI were reduced obviously after treatment as compared with those before treatment in two groups (all P < 0.05). The improvements in the Shu-needle therapy group were superior to those in the acupotomy group (both P < 0.05). In the follow-up observation, the recurrence rate in the Shu-needle therapy group was lower than that in the acupotomy group [17.8% (8/45) vs 46.4% (13/28), P < 0.05].

CONCLUSIONShu-needle therapy in combination with ozone injection of low concentration achieves the superior efficacy on lumbar disc herniation as compared with the acupotomy group.

Acupuncture Therapy ; Adult ; Combined Modality Therapy ; Female ; Humans ; Injections ; Intervertebral Disc Displacement ; drug therapy ; therapy ; Lumbar Vertebrae ; drug effects ; Male ; Middle Aged ; Ozone ; administration & dosage ; Treatment Outcome ; Young Adult

To investigate the effects of telmisartan on steatohepatitis (NASH) in rats by activating peroxisome proliferator-activated receptor gamma (r). Thirty male SD rats were randomized into normal control group, NASH control group and telmisartan prevention group. Normal control group was given standard food and the other two groups were given high fat diet for 16 weeks to induce NASH. Prevention group was given telmisartan (5 mg.kg-1.d-1) for 4 weeks by intragastric adminstration after 12 weeks. At the end of the 16th week, all the rats were sacrificed. Pathological changes of liver were observed by optical microscopy. Serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), fasting blood glucose(FBG), fasting insulin(FINS), HOMA-IR(homeostasis model assessment insulin resistance), Serum TNF-a and adiponectin were detected and analyzed.Western blot and RT-PCR were used to detect PPARr expression in hepatic tissues on protein and mRNA levels. (1) Rats were successfully modeled. The liver tissue samples were divided into 4 degrees (F0 - 4) based on total fatty degeneration of liver cells.There was one rat reached F3 and nine rats reached F4 in NASH group, one rat reached F1, six rats reached F2 and three rats reached F3 in prevention group. Inflammatory activity scores of hepatic tissues in the model group were 2.67+/-0.25, while that in the control group was 0 (U=15 and P is less than to 0.01), in the prevention group were 2.67+/-0.25 and 1.36+/-0.12 (U=24 and P is less than to 0.05 ). (2) The levels of serum ALT, AST, FBG, FINS, TNFa and HOMA-IR in the model group were increased than those in the control group( the vaules of q were 13.130, 6.472, 6.909, 26.619, 14.591 and 49.683 respectively, P less than 0.01). The levels of serum ALT, FINS, FBG, TNFa and HOMA-IR in the prevention group were decreased as compared to the model group (the vaules of q were 7.024, 4.145, 14.829, 13.195 and 31.991 respectively, P less than 0.01 ). (3) The serum adiponectin, PPARrmRNA and protein in liver tissues of the model group were lower than those in the control group (q values were 10.696, 8.679 and 16.762 respectively, P is less than to 0.05).The data in the prevention group were higher as compared to the model group(q values were 3.879,3.079,6.400, P is less than to 0.05 respectively). HOMA-IR was positively correlated with the expression of TNFa but negatively correlated with the expression of adiponectin (r = 0.927, P is less than to 0.01; r = -0.891, P is less than to 0.01, respectively). Telmisartan may has preventive effect on rats with steatohepatitis (NASH) by a mechanism of activating peroxisome proliferator-activated receptor r.
Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Fatty Liver ; Insulin Resistance ; Non-alcoholic Fatty Liver Disease ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the expression of PTEN and loss of heterozygosity (LOH) of its epigenetic microsatellite in gastric carcinoma and explore their roles in progression of gastric carcinoma.

METHODSLOH of epigenetic microsatellites of PTEN (D10S541, D10S583 and D10S1687) in advanced gastric cancer was detected by PCR-SSCP. Expression of PTEN mRNA and protein in normal gastric mucosa and gastric cancer was evaluated by RT-PCR and SABC immunohistochemistry, respectively. The relationship between expression of PTEN mRNA and protein and lymph node metastasis or LOH of microsatellites was discussed.

RESULTSLOH of D10S541, D10S583 and D10S1687 was found in 37.5% (21/56) of advanced gastric cancers. The positive rates of PTEN mRNA expression were 80.4% (45/56), 45.5% (5/11) and 32.1% (18/56) in normal mucosa, early and advanced gastric carcinomas, respectively, while 78.6% (44/56), 44.5% (5/11) and 28.6% (16/56) at the protein level. PTEN mRNA and protein were less frequently expressed in early and advanced gastric carcinomas than that in normal gastric mucosa (P < 0.05). There was positive correlation between PTEN mRNA expression and LOH of microsatellites in advanced gastric carcinomas. PTEN protein expression paralleled with its mRNA expression (P < 0.05). The expression of PTEN mRNA and protein was negatively correlated with lymph node metastasis of advanced gastric carcinomas (P < 0.05).

CONCLUSIONDown-regulated expression of PTEN gene is found in different stages of gastric carcinoma, and is closely correlated with LOH of its epigenetic microsatellites, which probably is its underlying molecular mechanisms. It suggests that altered PTEN gene contributes to tumorigenesis and progression of gastric carcinomas.

Gastric Mucosa ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Loss of Heterozygosity ; Lymphatic Metastasis ; genetics ; Microsatellite Repeats ; genetics ; Neoplasm Staging ; PTEN Phosphohydrolase ; Phosphoric Monoester Hydrolases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Tumor Suppressor Proteins ; biosynthesis ; genetics