Objective To investigate a method for the purification of the N?terminal peptide fragment(NT)of the myocardial calcium channel Cav1.2,and characterize its interaction with calmodulin(CaM). Methods EscherichiacoliBL?21 cells were transformed with plasmid pGEX?6p?3/NT harboring the NT?GST fusion gene. The cells harboring pGEX?6p?3/NT were cultured and protein expression was induced with isopropyl?β?D?thiogalactoside(IPTG). Then,the GST?NT fusion protein was purified by using glutathione Sepharose 4B(GS?4B)beads. GST was cleaved off with the PreScission protease,and SDS?PAGE was performed to detect the purity and relative molecular weight of the purified peptide. Further, GST pull?down assay was performed to characterize the interaction of the NT peptide with CaM. Results SDS?PAGE analysis showed that the NT peptide was successfully purified,with high purity. Results of the GST pull?down assay showed that the NT peptide could interact with CaM. Conclusion This study establishes a method for the purification of the NT peptide and lays the foundation for further research on the interaction partners and biological functions of NT.

Objective To study the onset features and axiom of children disease in Duanguan region.Methods A statistic study was carried on 1 782 cases of children disease which confirmed by pathological examinations.The sex,age and pathology(congenital malformation,inflammatory disease,tumor-like lesions,benign tumors,malignant tumors)were registered.Results The inflammatory disease occupied the first place in incidence rate(44.61%),followed by tumor-like lesions(25.03%),benign tumors(23.34%),congenital malformation(5.22%)and malignant tumors(1.80%).Among inflammatory disease,appendicitis was the most common,the next was chronic tonsillitis and tuberculosis.Among tumor-like lesions,cystic disease was the most common;the next was polyps.Among benign tumors,vasculoma was the most common,the next was pilomatricoma,osteoid and chonodroid tumors.Among congenital malformation,ear fistula was the most common,the next was diverticulosis and intestineal duplicateion.Among the malignant tumors,the sarcoma of soft tissue was the most common,the next was the malignant peripheral nerve tumors and lymphoma.Conclusion Children's disease have its own characteristics which varied in age,sex and pathological types,and differ from those of adults.

BACKGROUNDThe result would be disastrous if the superior pulmonary sulcus tumor (Pancoast tumor) was misdiagnosed as degenerative cervical spine diseases. The aim of this study was to investigate the differential diagnosis methods of cervical radiculopathy and superior pulmonary sulcus tumor.

METHODSClinical manifestations, physical, and radiological findings of 10 patients, whose main complaints were radiating shoulder and arm pain and later were diagnosed with superior pulmonary sulcus tumor, were reviewed and compared with those of cervical radiculopathy.

RESULTSSuperior pulmonary sulcus tumor patients have shorter mean history and fewer complaints of neck pain or limitation of neck movement. Physical examination showed almost normal cervical spine range of motion. Spurling's neck compression test was negative in all patients. Anteroposterior cervical radiographs showed the lack of pulmonary air at the top of the affected lung in all cases and first rib encroachment in one case. The diagnosis of superior pulmonary sulcus tumor can be further confirmed by CT and MRI.

CONCLUSIONSBy the method of combination of history, physical examination, and radiological findings, superior pulmonary sulcus tumor can be efficiently differentiated from cervical radiculopathy. Normal motion range of the cervical spine, negative Spurling's neck compression test, and the lack of pulmonary air at the top of the affected lung in anteroposterior cervical radiographs should be considered as indications for further chest radiograph examinations.

Adult ; Aged ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Pancoast Syndrome ; diagnosis ; Radiculopathy ; diagnosis

BACKGROUNDThe association between vulnerability of plaque assessed with intravascular ultrasound (IVUS) and plasma levels of fibrinolytic biomarkers was determined in patients with acute coronary syndrome (ACS). However, few data are available on the relationship between the levels of tissue type plasminogen activator (t-PA) and virtual histological intravascular ultrasound (VH-IVUS) signs of plaque instability.

METHODSEighty-nine patients with ACS were enrolled in the study. Blood was collected to measure t-PA levels by liquid phase bead flow cytometry. Eighty-nine nonbifurcate lesions (identified by coronary angiography and ECG) were investigated using IVUS before catheterization. IVUS radiofrequency data obtained with a 20 MHz catheter were analyzed with IVUS virtual histological software. The areas of plaque and media were calculated and lesions were classified into two groups: VH-IVUS derived thin cap fibroatheroma (VH-TCFA) and non-VH-TCFA plaque.

RESULTSPlasma t-PA level in the patients with TCFA was significantly lower than that with non-TCFA ((1489+/-715) pg/ml vs (2163+/-1004) pg/ml). Decreased plasma levels of t-PA were associated with plaque vulnerability. Plasma levels of t-PA correlated negatively with plaque plus media and necrotic core in plaque in patients with ACS.

CONCLUSIONSt-PA is an independent risk factor and a powerful predictor of vulnerable plaques. Decreased levels of t-PA may reflect instability of atherosclerotic plaques and might therefore serve as noninvasive determinants of those at high risk for consequent adverse events.

Acute Coronary Syndrome ; blood ; pathology ; Aged ; Coronary Artery Disease ; pathology ; Coronary Vessels ; pathology ; Female ; Humans ; Male ; Middle Aged ; Tissue Plasminogen Activator ; blood ; Ultrasonography, Interventional

OBJECTIVETo investigate whether WASP/Verprolin homologous protein 1 (WAVE1) plays a role in the pathogenesis of childhood acute lymphoblastic leukemia (ALL).

METHODSWAVE1 mRNA and protein expression in bone marrow mononuclear cells (BMMCs) was measured by RT-PCR and Western blotting respectively in 4 children with ALL relapse, 15 children with ALL in complete remission (CR) and 40 children with newly diagnosed ALL. Ten normal bone marrow samples were used as controls. Jurkat cells were treated with different concentrations of adriamycin (ADM). The cell proliferation was detected with MTT. The apoptosis rate was measured by flow cytometry. WAVE1 mRNA and protein expression of Jurkat cells treated with ADM was detected by RT-PCR and Western blotting respectively.

RESULTSWAVE1 was not expressed or weakly expressed in BMMCs from normal controls and patients with ALL in CR. Higher WAVE1 mRNA and protein expression was found in BMMCs from patients with newly diagnosed ALL and patients with relapse ALL when compared with the controls and the patients in CR (P<0.01). ADM significantly inhibited the proliferation of the Jurkat cells and the inhibitory effect was dose-and time-dependent (P<0.05). After ADM treatment for 24 hrs, the percentage of apoptosis cells increased significantly and WAVE1 mRNA and protein expression of Jurkat cells decreased significantly when compared with the untreated controls (P<0.05).

CONCLUSIONSThe WAVE1 expression increased in children with ALL. WAVE1 may be related to the development of ALL and may be severed as a marker for the evaluation of the severity of ALL in children.

Adolescent ; Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Cell Proliferation ; drug effects ; Child ; Child, Preschool ; Doxorubicin ; pharmacology ; Female ; Humans ; Infant ; Jurkat Cells ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; etiology ; metabolism ; RNA, Messenger ; analysis ; Wiskott-Aldrich Syndrome Protein Family ; analysis ; genetics ; physiology

OBJECTIVETo investigate the feasibility, effectiveness and practicability of transurethral enucleation plus pneumocystostomy rotary cut (TUE + PCRC) for large benign prostatic hyperplasia (BPH).

METHODSWe performed TUE + PCRC for 26 BPH patients aged 62 - 85 years with the prostate volume of 80 - 165 ml. We conducted transurethral enucleation of the hyperplastic prostate glands and pushed them into the bladder, followed by bladder puncture for pneumo-cystostomy rotary cut.

RESULTSAll the surgical procedures were successfully accomplished, with the mean surgical time of 41 (32 - 54) minutes and intraoperative blood loss < 60 ml in all the cases. Twenty-three of the patients were followed up for 2 - 8 months, which revealed no stricture of the urethra or any other severe complications. Compared with the preoperative baseline, significant improvement was achieved in the IPSS (6.5 +/- 2.2 vs 26.2 +/- 2.4), QOL (1.4 +/- 0.9 vs 4.6 +/- 1.2) and Qmax ([5.8 +/- 1.0 ] vs [19.6 +/- 2.8] ml/s) of the patients after surgery (P < 0.01).

CONCLUSIONTUE + PCRC, with its advantages of short operation time and less severe complications, is a safe and effective approach to the management of large BPH.

Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Prostatic Hyperplasia ; surgery ; Transurethral Resection of Prostate ; methods

OBJECTIVETo provide scientific basis for the selection of agrotype and property fertilization for Rumex gmelini cultivated in compliance with good agricultural practice (GAP).

METHODHPLC method was applied to determinate the content of seven active constituents (resveratrol, polydatin, chrysophanol 1-glucoside, nepodin, emodin, chrysophanol and physcion) of annual R. gmelini. And the correlation between soil nutrients and content of active constituents in the root of R. gmelini were analyzed by stepwise regression analysis.

RESULTSeven regression equation were established. The statistic significance was found in three of them.

CONCLUSIONThe soil with high total K level is not suitable for R. gmelini cultivation. But the higher available N, available P, available K level of soil is suitable.

Anthraquinones ; analysis ; Ecosystem ; Emodin ; analogs & derivatives ; analysis ; Fertilizers ; Glucosides ; analysis ; Nitrogen ; analysis ; Phosphorus ; analysis ; Plant Roots ; chemistry ; growth & development ; Plants, Medicinal ; chemistry ; growth & development ; Potassium ; analysis ; Regression Analysis ; Rumex ; chemistry ; growth & development ; Soil ; analysis ; Stilbenes ; analysis

OBJECTIVETo establish a method used for optimization of harvesting time and determine the best time for harvesting Rumex gmelini.

METHODAn HPLC method was applied to determinate the contents of seven active constituents(resveratrol, polydatin, chrysophanol 1-glucoside, nepodin, emodin, chrysophanol and physcion)of R. gmelini at different development stage. The result was analyzed by principal component analysis.

RESULTThe accumulation of active constituents showed a regular pattern.

CONCLUSIONThe best harvesting time of R. gmelini is early July.

Anthraquinones ; analysis ; Emodin ; analogs & derivatives ; analysis ; Glucosides ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; growth & development ; Principal Component Analysis ; methods ; Quality Control ; Rumex ; chemistry ; growth & development ; Seasons ; Stilbenes ; analysis

BACKGROUNDWeight gain following smoking cessation increases cardiovascular risk, but its effects on prognosis after percutaneous coronary intervention (PCI) remain unclear. This study aimed to investigate the relationship between weight gain post smoking cessation and one-year clinical outcome in patients who underwent PCI with drug-eluting stent (DES).

METHODSA total of 895 consecutive male smoking patients were divided into quitters (n = 437) and continuers (n = 458) according to their smoking status after PCI. Weight gain, major adverse cardiac events (MACE, including cardiac deaths, myocardial infarction and revascularization), and recurrent angina were recorded during follow-up for one year.

RESULTSAverage weight gain in quitters was more than that in continuers (1.5 kg vs. -0.03 kg, P < 0.001). Weight was unchanged or increased by more than 1.5 kg in 78.17% of continuers, while 50.57% of quitters had a weight gain of less than 1.5 kg. Compared with continuers, MACE in quitters was significantly reduced after PCI (6.12% vs. 4.81%, P = 0.049), especially recurrent angina (13.97% in continuers vs. 9.84% in quitters, P = 0.027). After adjusting for weight gain and other factors, smoking cessation was independently associated with a lower risk of MACE and recurrent angina (OR = 0.73, P = 0.035). However, weight gain > 1.5 kg (OR = 1.55, P = 0.026) could curtail the benefits from smoking cessation.

CONCLUSIONSWeight gain may reduce the benefits of smoking cessation after PCI with DES implantation. Thus, although smoking cessation is recommended after PCI, weight control should also be highly encouraged for these patients.

Aged ; Angioplasty, Balloon, Coronary ; Drug-Eluting Stents ; Humans ; Middle Aged ; Smoking Cessation ; Weight Gain

OBJECTIVETo analyze the postoperative short-term and long-term outcomes in the management of type I esophageal atresia, and to explore the ideal operative strategy.

METHODSClinical data of 22 patients with type I esophageal atresia treated from January 2005 to September 2012 were retrospectively reviewed. Of 22 patients, 6 patients gave up the treatment. Two underwent primary repair after birth. Of 14 patients undergoing cervical esophagostomy and gastrostomy, 8 patients received esophageal replacement. Postoperative short-term and long-term complications, nutritional state and neurodevelopment were studied on above 10 children with radical operations.

RESULTSOf 10 patients with radical operation, the short-term complications were hydrothorax in 1 case, anastomotic leakage in 4, dumping syndrome in 1, anastomotic stricture in 1. The long-term complications were esophageal stricture in 2 cases, and repeated respiratory infection in 3. These complications could be managed successfully. The postoperative follow-up duration ranged from 2 to 62 months. Two cases were lost during follow-up after 2 years. Weight-for-age was normal in 2 patients, mild malnutrition in 5 patients, and moderate malnutrition in 1 patients. Neurodevelopment is significantly delayed as compared to normal children.

CONCLUSIONSOperative strategy should be chosen according to the distance between proximal and distal esophagus in the treatment of type I esophageal atresia. The efficacy of radical operation is relative satisfactory in terms of short-term and long-term complications and the quality of life.

Child ; Esophageal Atresia ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Postoperative Complications ; Quality of Life ; Retrospective Studies