AIM To study the effect of genistein (GST), a protein tyrosine kinases inhibitor, on L-type calcium channel currents (Iba,L or Ica,L, dependent on permeating ion used) in freshly dispersed colon smooth muscle cells from guinea-pig. METHODS Single colon smooth muscle cells were enzymatically dissociated from guinea-pig. L-type calcium currents were measured by conventional whole-cell patch-clamp techniques. RESULTS The peak amplitudes of Iba,L elicited to 10 mV test potential from a holding potential of -80 mV, were reversibly and dose-dependently reduced by GST (10-100 μmol·L-1) with an IC50 value of (39.9±3.6)μmol·L-1. Bath application of GST shifted the steady-state inactivation curves of Iba,L in a hyperpolarized direction (about 10 mV, P＜0.01) without altering their slopes. The peak amplitudes of Iba,L were also inhibited but to a less extent by daidzein, an inactive analogue of GST. Sodium orthovanadate 1 mmol·L-1, a potent inhibitor of protein tyrosine phosphatases, blocked GST-induced inhibition of Ica,L. CONCLUSION GST can block L-type calcium channel activity in guinea-pig colon smooth muscle cells via tyrosine kinase pathway.

To investigate the characteristics of large-conductance calcium-activated potassium channels (BK(Ca)) and the effect of hydrogen peroxide (H2O2) on BK(Ca) in guinea-pig proximal colon smooth muscle cells, single smooth muscle cells of guinea-pig colon were enzymatically isolated in low calcium solution containing papain (3 mg/mL), DTT (2 mg/mL), and bovine serum albumin (BSA, 2 mg/mL). Tissues were incubated at 36 degrees C in enzyme solution for 15 min and were then suspended in enzyme-free low calcium solution. Inside-out single channel recording technique was used to record BK(Ca) current. The intracellular (bath) and microelectrode solution both contained symmetrical high potassium. The BK(Ca) in guinea-pig colon smooth muscle cell possesses: 1) voltage-dependence, 2) high selectivity for potassium ion, 3) large conductance (223.7 pS+/-9.2 pS), 4) dependence of [Ca(2+)](i). Intracellular application of H2O2 decreased the open probability (P(o)) of BK(Ca) at low concentration ( Animals ; Colon ; cytology ; Guinea Pigs ; Hydrogen Peroxide ; pharmacology ; Large-Conductance Calcium-Activated Potassium Channels ; physiology ; Myocytes, Smooth Muscle ; physiology ; Oxidation-Reduction

Eleven compounds were isolated and purified from the barks extract of Nothopanax delavayi and their structures were identified as serratagenic acid-3-O-alpha-L-arabinopyranosyl-28-O-beta-D-glucopyranosyl ester (1), serratagenic acid-3-0-alpha-L-arabi-nopyranosyl-28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester (2), serratagenic acid (3), serratagenic acid-3-O-alpha-L-arabinopyranoside (4), serratagenic acid-beta-O-beta-(2', 4'-O-diacetyl) -D-xylopyranosyl-28-O-[alpha-L-rhamnopy-ranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->46)-beta-D-glucopyranosyl] ester (5), serratagenic acid-3-O-alpha-(4'-O-acetyl)-L-arabino pyrano-syl-28-0- [-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester(6), serratagenic acid-3-O-alpha-(2'-O-acetyl)-L-arabinopyranosyl-28-O-[-alpha-L-rhamnopyranosyl- (1-->4) -beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester(7), serratagenic acid-3-0-beta-D-xylopyranosyl-28-O-[-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl] ester (8), protocatechuic acid (9), ethyl caffeate (10) and caffeic anhydride (11) by physicochemical properties and spectroscopic data analysis. Among them, compounds 3-4 and 9-11 were firstly isolated from the genus Nothopanax, and compounds 5-8 were isolated from this plant for the first time.
Araliaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Bark ; chemistry

The aim of the present study was to investigate the effects of quercetin on colon contractility and voltage-dependent Ca(2+) channels in the single smooth muscle cell isolated from the proximal colon of guinea-pig and to clarify whether its effect on L-type Ca(2+) current (I(Ca,L)) would be related to its myorelaxing properties. Colon smooth muscle strips were used to take contractile tension recordings. Smooth muscle cells were freshly isolated from the proximal colon of guinea-pig by means of papain treatment. I(Ba,L) (barium instead of calcium as current carrier) was measured by using whole-cell patch-clamp techniques. The results showed that quercetin relaxed colon muscle strips in a concentration-dependent manner and antagonized the contractile effect of acetylcholine and neostigmine. Preincubation with indomethcin [cyclooxygenase (COX) inhibitor] and methylene blue [guanylate cyclase (GC) inhibitor] significantly attenuated the relaxing effect of quercetin, respectively. Quercetin increased I(Ba,L) in a concentration- [EC(50)= (7.59+/-0.38) mumol/L] and voltage-dependent pattern, and shifted the maximum of the current-voltage curve by 10 mV in the depolarizing direction without modifying the threshold potential for Ca(2+) influx. Quercetin shifted the steady-state inactivation curve toward more positive potentials by approximately 3.75 mV without affecting the slope of activation and inactivation curve. H-89 (PKA inhibitor) abolished quercetin-induced I(Ba,L) increase, while cAMP enhanced the quercetin-induced I(Ba,L) increase. The patch-clamp results proved that quercetin increased I(Ba,L) via PKA pathway. It is therefore suggested that the relaxing effect of quercetin attributes to the interaction of GC and COX stimulation, as well as the antagonism effect on acetylcholine, which hierarchically prevails over the increase in the Ca(2+) influx to be expected from I(Ca,L) stimulation.
Animals ; Calcium Channels, L-Type ; metabolism ; Cells, Cultured ; Colon ; drug effects ; Guinea Pigs ; Muscle Contraction ; Myocytes, Smooth Muscle ; drug effects ; Patch-Clamp Techniques ; Quercetin ; pharmacology

Objective:To observe the effect of moxibustion on the mRNA and protein expressions of heat-shock protein 70 (HSP70) in gastric cancer-bearing rats. Methods: A total of 40 healthy Sprague-Dawley (SD) rats were adaptively fed for one week. The gastric cancer model was prepared by Walker-256 cancer tissue transplantation. After 7 d, 10 rats were randomly selected to verify the successful modeling, and the remaining 30 rats were divided into a model group, a moxibustion group and an infrared group by the random number table method, with 10 rats in each group. After enrollment, the moxibustion group received suspended moxibustion at Zhongwan (CV 12), Guanyuan (CV 4) and bilateral Zusanli (ST 36), (the first group of acupoints) on the 1st day, and suspended moxibustion at bilateral Pishu (BL 20) and Weishu (BL 21), (the second group of acupoints) on the 2nd day, 20 min each time, once a day. Moxibustion was alternately performed every other day at the two groups of acupoints for 21 d. From the day of enrollment, rats in the infrared group were irradiated with the infrared radiation at the stomach area on the 1st day, and at the T12-T13 interspinous region on the 2nd day, 20 min each time, once a day, and the two locations were alternately irradiated every other day for 21 d. During the treatment, rats in the model group were intervened by grasping and fixation without treatment. At the end of the treatment, blood was collected from the inner eye orbit, and the HSP70 expression in peripheral blood was determined by enzyme linked immunosorbent assay (ELISA). Rats were sacrificed, the tumor volume and growth inhibition rate were measured. The position and changes of HSP70 in gastric cancer were observed by streptavidin-perosidase (SP); HSP70 protein expression was determined by ELISA; HSP70 mRNA expression in cancer tissues was determined by reverse transcription-polymerase chain reaction (RT-PCR) assay. Results: In comparison of the model group, the volume growth of the gastric cancer in the moxibustion group was significantly restricted (P＜0.01); the volume growth inhibition rate in the moxibustion group was 37.93%; the HSP70 expression in peripheral blood and the cancer tissues was significantly increased (both P＜0.01); the expression of HSP70 mRNA and HSP70 content in gastric tumor were both obviously increased in the moxibustion group (P＜0.01); and a large amount of HSP70 was released to the outside of cancer cells in the moxibustion group. In comparison of the model group, the volume growth of the gastric cancer in the infrared group was slightly restricted (P＜0.05) with a volume growth inhibition rate of 15.89%; the HSP70 expression in the infrared group was increased significantly in peripheral blood (P＜0.01) and in the gastric cancer tissues (P＜0.05); more HSP70 was released outside of the cancer cells in the infrared group. In comparison of the infrared group, the volume growth of gastric cancer was more restricted in the moxibustion group (P＜0.05), and the HSP70 expression in the gastric cancer tissues was also higher (P＜0.05); more HSP70 was released outside of the cancer cells in the moxibustion group. Conclusion: Moxibustion and infrared treatment inhibit the gastric cancer growth in the gastric cancer-bearing rats, up-regulate the HSP70 expression in gastric cancer tissues, and promote the production and extracellular release of HSP70, and the effect of moxibustion is more obvious.

OBJECTIVETo observe the efficacy of hybrid balloon valvuloplasty for the treatment of low-body weight infants with severe congenital valvular aortic stenosis (AS).

METHODSFive infants with severe congenital valvular aortic stenosis underwent the hybrid balloon aortic valvuloplasty through median sternotomy in the hybrid operating room. The mean age was (40.2 ± 7.0) days, weight was (4.48 ± 0.75) kg. The patients were followed up by echocardiography for 9 - 13 months post procedure.

RESULTSOperation was successful in all 5 patients and they were discharged from hospital uneventfully. The gradient pressure decreased significantly from (98.8 ± 9.0) mm Hg (1 mm Hg = 0.133 kPa) to (13.8 ± 3.3) mm Hg (P < 0.05) post operation. There was no moderate or severe aortic insufficiency. All patients were alive, the gradient pressures was (18.8 ± 2.5) mm Hg and there was no moderate or severe aortic insufficiency during follow-up [9 - 13 (11.0 ± 1.4) months].

CONCLUSIONThe hybrid balloon aortic valvuloplasty is an effective option for the low-body weight infants with severe congenital valvular aortic stenosis.

Aortic Valve Stenosis ; congenital ; surgery ; Balloon Valvuloplasty ; Female ; Humans ; Infant ; Infant, Low Birth Weight ; Infant, Newborn ; Male

OBJECTIVE: To investigate the early diagnosis of breast cancer with nonpalpable tumor. METHODS: Forty-six cases of clinical nonpalpable tumor were examined by mammography and sonography.The cases of nipple discharge were also examined by fiberoptic ductoscopy. RESULTS: Breast cancer in 46 cases was diagnosed by pathological examination. Of them, 34 diagnosed with breast cancer were found with nodus, calcification or confused structure, and so on, 5 were considered benign tumor, and 7 were not found lesion by mammography. Thirty-one cases were diagnosed with breast cancer, 6 with benign tumor, and 9 were not found occupying lesion by sonography. Occupying lesions were found in 6 cases of nipple discharge by fiberoptic ductoscopy and were finally diagnosed by biopsy. CONCLUSION Mammography and sonography are important methods in early discovering breast cancer. Early diagnosis rate of breast cancer can be elevated by signs of early breast cancer and combined examination of mammograply and sonography.
Adult ; Aged ; Breast Neoplasms ; diagnosis ; pathology ; Carcinoma, Ductal, Breast ; diagnosis ; pathology ; Female ; Humans ; Mammography ; Middle Aged ; Ultrasonography, Mammary

Objective To compare the difference in pulse pressure variation(PPV)to predict volumetric response in children with ventricular septal defect(VSD)and tetralogy of Fallot(TOF). Methods VSD group consisted of 38 patients,aged 1.05 ± 0.75 years,while TOF group consisted of 36 patients,aged 1.15 ± 0.68 years. After separation from cardiopulmonary bypass,fluid infusion therapy was administered. PPV was recorded using pressure recording analytical method along with cardiac index(CI)before and after fluid infusion. Patients were considered as responders to fluid loading when CI increased ≥15%.Receiver operating characteristic(ROC) analysis was used to assess the accuracy and cutoffs of PPV to predict volumetric response.Results The PPV val-ues before and after fluid infusion were significantly lower in TOF group than that in VSD group(P < 0.01 for both).In VSD group,27 were responders and 11 nonresponders.ROC curve area was 0.89 and cutoff value 17.4%. In TOF group,26 were responders and 10 nonresponders.ROC curve area was 0.79 and cutoff value 13.4%.Con-clusion PPV is predictive of volumetric response in VSD and TOF patients following cardiac surgery.PPV's pre-dictivity and cutoff value are higher than the former. PPV is affected by right ventricle-pulmonary artery circula-tion,under-fluid infusion should be avoided in TOF due to lower PPV,over-fluid infusion should be avoided in VSD due to higher PPV.

Carbon nanosheets load beta-cyclodextrin (β-CD-CNS) as a new modified electrode materials was reported for the electrochemical determination of sulfadiazine(SD). Carbon nanosheets(CNS) were prepared by a new method of ultrasonic electrolysis in which the β-CD was attached on CNS through ultrasonic dispersion method. The β-CD-CNS composite nanomaterials were immobilized onto glassy carbon electrodes with drops of coating method to construct an SD voltammetric sensor. The differential pulse stripping voltammetry (DPSV) was used to characterize the electrocatalytic behavior of the developed sensor. The Effects of some parameters on the response behavior of the sensor such as pH,modification amount,scanning rate,stirring speed,stirring time,deposition potential and time were investigated and optimized. The results indicated that the β-CD-CNS composite nanomaterials had excellent electroactivity for the SD in neutral solution and greatly improved the current response of SD. Under the optimal conditions, the SD had an irreversible characteristic oxidation peak around+0.87 V,and the oxidation peak current ip(μA) had a good linear relationship with the concentration C ( μmol/L) of the SD in concentration range of 0.05 μmol/L-13.5 μmol/L with correlation coefficients of 0.999. The detection limit was 12.2 nmol/L (S/N=3). The sensor was successfully applied for the trace SD determination in water and milk samples and the recoveries from the spiked samples were 80.0%-102% with RSD≤5.2%.

OBJECTIVETo determine the clinical value of 24 h double-probe pH-metry for the diagnosis and treatment of laryngopharyngeal reflux (LPR).

METHODSAccording to the pH-metry results (whether the reflux events record in the upper esophagus is more than 6.9), patients of refractory pharyngolaryngitis were divided into LPR and control groups (each of 17 cases). All patients treated with anti-acid therapy and conventional pharyngo-laryngitis therapy. Correlation between pH-metry and the reflux symptom index (RSI), the reflux symptom index (RFI) were analyzed. Changes of the RSI and RFI in different group were calculated in post-treatment.

RESULTSIn the LPR group, the median reflux events of the upright time were higher than the supine time (Z = -3.62, P < 0.01), but the difference was not discovered in the control group (Z = -0.60, P > 0.05). There was no statistical difference between RSI, RSI and RFS with pH-metry, and with moderate concordance (k were 0.47, 0.53, P < 0.01, respectively). Compared to pre-treatment, the RSI and RFI were decreased both in LPR group and control group, Amplitude decreased in LPR group significantly higher than the control groups, with statistical difference (t were 3.74, 3.01, P < 0.01, respectively).

CONCLUSIONThe 24 h double-probe pH-metry is significant for the anti-acid therapy of LPR.

Adult ; Esophageal pH Monitoring ; Female ; Gastroesophageal Reflux ; diagnosis ; Humans ; Laryngopharyngeal Reflux ; diagnosis ; Male ; Middle Aged