To approach the method of isolation of tolerant human atrial myocytes, single myocytes were isolated by modified procedure of enzymatic dissociation with protease (type XXIV) and collagenase (type V). L-type calcium channel current (I(Ca-L)), sodium current (I(Na)), transient outward potassium current (I(to1)), and inward rectifier potassium current (I(K1)) in isolated atrial myocytes were recorded by using whole-cell patch-clamp techniques. Single cardiocytes isolated by this method were smooth, well-striated and rod-shaped. The yields of recordable myocytes, which viable and calcium-tolerant for electrophysiological studies, were 50%-60% of the total isolated cells. Compared with other isolation methods, this method was simple and steady, but with yield of a great number of qualified myocytes. The currents recorded in these cells were functional and active. Our research suggests that the myocytes isolated by the described method in this paper have normal electrophysiological function and are appropriate for patch-clamp experiments.
Cell Separation ; methods ; Humans ; Myocardium ; cytology ; Myocytes, Cardiac ; cytology ; Patch-Clamp Techniques

Objective To explore the influence of parental history of allergy on the cytokine level of umbilical cord blood of neonates.Methods A prospective study was conducted which selected the pregnant women attending Yiwu maternal and child healthcare hospital from July to December in 2012 as objects.The umbilical cord blood sample were collected to detect the level of IgE,eosinophile granulocyte,eotaxin,IL -9,IL -6,IL -4,IL -5,IFN -γand IL -10. Results A total of 136 subjects were recruited and divided into three groups according to the status of parental allergy histories.There was no significant difference in social and demographic characteristics among the three groups (P>0.05). Three groups have significantly different median concentration of IL-13 (1.82pg/mL vs.3.24pg/mL vs.6.12pg/mL), eotaxin (29.33pg/mL vs.50.71pg/mL vs.60.15pg/mL),IL-9 (43.75pg/mL vs.111.17pg/mL vs.183.19pg/mL), IL-6 (11.49pg/mL vs.19.35pg/mL vs.26.09pg/mL),IL-4 (3.67pg/mL vs.21.27pg/mL vs.49.51pg/mL),IL-5 (4.26pg/mL vs.7.69pg/mL vs.10.66pg/mL),IFN-γ(338.65pg/mL vs.649.17pg/mL vs.834.14pg/mL),IL-10 (7.13pg/mL vs.10.56pg/mL vs.14.64pg/mL),P <0.05.Conclusion Parental history of allergy could have influence on cytokine level of umbilical cord blood.Early screening and surveillance in neonates whose parents have history of allergy should be taken to guide early interventions.

Microsatellite instability-high (MSI-H) colorectal cancer accounts for approximately 10%-15% of all colorectal cancer patients, while in metastatic diseases the MSI-H population accounts for only 5% of patients. Previous studies have shown that early-stage MSI-H colorectal cancer patients have a good prognosis, but those with advanced disease have a poor prognosis and are not sensitive to chemotherapy. The advent of PD-1 antibodies has significantly improved the prognosis and changed treatment landscape in this population, not only achieving good outcomes in late-line therapy, but also significantly outperforming traditional chemotherapy combined with targeted therapy in first-line therapy. How to overcome primary and secondary drug resistance is a key issue in improving the outcome of MSI-H metastatic colorectal cancer, and commonly used approaches include changing chemotherapy regimens, combining with other immunotherapies, combining with anti-angiogenesis, and local treatments (surgery, radiotherapy, or interventional therapy). It is worth noting that immunotherapy has certain lifelong or even lethal toxicity, and the indications for neoadjuvant immunotherapy must be evaluated with caution. Neoadjuvant immunotherapy in MSI-H advantaged population can achieve high rates of pathological complete remission (pCR) and clinical complete remission (cCR). Therefore, for MSI-H patients with a strong intention to preserve anal sphincter and a strict evaluation of cCR after neoadjuvant immunotherapy, the Watch-and-Wait strategy offers an opportunity to preserve sphincter function and improve long-term survival quality in a subset of mid-to-low rectal cancers. Research on adjuvant immunotherapy in the field of colorectal cancer is also in full swing, and the results are worth waiting for.
Colonic Neoplasms ; Colorectal Neoplasms/therapy* ; Humans ; Immunotherapy/methods* ; Microsatellite Instability ; Microsatellite Repeats

OBJECTIVEThe aim of the study was to determine the prevalence and the distribution pattern of lesion site of intracranial vascular stenosis and to identify risk factors for the stenosis in patients with essential hypertension.

METHODSA total of 231 consecutive inpatients with essential hypertension were included in this study. Patients with the history of cerebrovascular diseases and relevant neurological symptoms were excluded. Intracranial vascular stenosis (>50% diameter reduction) was detected using CT angiography (CTA).

RESULTSOf 231 patients, 69 (29.87%) had intracranial artery stenosis. The most common stenosis site is middle cerebral artery (43.69%), followed by carotid siphon (20.39%). The stenosis in internal carotid arterial system (78.64%) was more common than in vertebrobasilar arterial system (21.56%, P < 0.05). The patients with intracranial vascular stenosis were older, had longer history of hypertension, higher levels of systolic blood pressure, higher plasma cholesterol, higher LDL-C. Lp (a), higher urinary microalbumin excretion, thicker ventricular septum, and lower levels of HDL-C than the patients without stenosis. Logistic analysis showed that systolic blood pressure (OR 1.650, 95% CI 1.134 - 2.400, P = 0.023), course of hypertension (OR 1.238, 95% CI 1.072 - 1.429, P = 0.006), LDL-C (OR 2.103, 95% CI 1.157 - 3.823, P = 0.014) and type 2 diabetes (OR 2.325, 95% CI 1.161 - 4.341, P = 0.011) were the independent risk factors of asymptomatic intracranial arterial stenosis.

CONCLUSIONSNearly 30% inpatients with essential hypertension had asymptomatic intracranial artery stenosis. The most common site of stenosis was middle cerebral artery. Hypertension, dyslipidemia and diabetes were risk factors for the development of intracranial arterial stenosis.

Aged ; Cerebral Angiography ; Female ; Humans ; Hypertension ; epidemiology ; pathology ; Intracranial Arterial Diseases ; epidemiology ; Male ; Middle Aged ; Prevalence ; Risk Factors

Objective To investigate epidemical situation of drinking-tea fluomsis in Qinghai province, in order to understand diet structure to provide the scientific basis for the prevention and control and the scientific research. Methods In 2007, according to "Scheme for Survey on Epidemical Drinking-tea Fluorosis", we carried out an customized investigation in 28 counties, 3 townships sampled in each county, 2 villages in each township, 50 adults and 50 school-age children in each village; at same time, 1 monk temple was sampled in each county, 50 clergy adults and 50 children in each temple. Then we investigated the resident income, the diet structure, the brick tea consumption and so on, and applied Dean method to diagnose dental fluorosis. The patient number estimated based on the survey result. Results ①Yeady per capita income of people was mostly 500 - < 1000yuan, next 1000 - < 3000 yuan; economic income in pasturing area was higher than that of agriculture, half area and half agriculture and half pastoral region and township. ②Staple food was bread flour primarily in the animal husbandry agricultural half pastoral area,next were the roasted barley and the rice;the bread flour was the principle food in the agricultural region and the cities,next were the rice and the roasted barley;among non-staple food,meat came fimt and milk foHowed,egg the last.③The frequently edible vegetables Was potato.cabbage and greenpepper,eaten by a majority of people[62.82%(6497/10 343)];as for fruits,apple,pear and orange was primarily consumed,75.95%(7856/10 343)of people ate less than 10 times every month.④Sixty-one thousand nine hundred and ninety-nine residents were registered,153 335 kg ofbrick tea was consumed in villages and towns,2.47 kg per person;in 1001 monks investigated,4120 kg of brick tea was consumed every year,4.12 kg per person.⑤Detection rate of adult dental fluorosis in the villages and towns was 24.11%(2494/10 343),that of the children was 24.38% (3012/12 355);detection rate of dental fluorosis in monks was 26.13%(203/777),that of the children was 39.73% (89/224).⑥Detection rate of adult skeletal fluorosis in villages and towns was 15.60%(17/109);that of monks was 4.88%(2/41).⑦The 95%confidence limit estimated a total number of dental fluorosis patients were 1 084 306- 1 134 170 persons.the median Was 1 109 238 persons;the 95% confidence limit estimated a total number of skeletal fluorosis patients were 309 177-758 199 persons,the median was 533 688 persons.Conclusions Qinghai province has a great quantity of brick tea consumption,having lots of people with drinking-tea fluorosis which is in severe degree.The resident food structure was monotonous and mostly transpolted from other region.

OBJECTIVETo investigate the changes of L-type Ca(2+) channel current (I(Ca-L)) and its voltage-dependent activation and inactivation in atrial myocytes of patients with atrial fibrillation (AF).

METHODSThe specimens of right atrial appendage were obtained from 18 patients with normal sinus rhythm (NSR) and 12 patients with chronic atrial fibrillation (CAF). Single myocytes were isolated by enzymatic dissociation with two-step method and the ionic currents were recorded using whole-cell patch clamp techniques to detect the changes of I(Ca-L) density and kinetic properties.

RESULTS(1) I(Ca-L) density was (-1.32 +/- 0.19) pA/pF in CAF group (n = 12) and (-4.58 +/- 0.39) pA/pF in NSR group (n = 21) at the test potential from -40 mV to 0 mV. I(Ca-L) density of CAF group was significantly reduced (P < 0.01), compared with the NSR group. (2) No significant differences were noted between the two groups in the voltage-dependent activation parameters (V(1/2), K) and inactivation parameters (V(1/2), K).

CONCLUSIONSI(Ca-L) density of CAF group was significantly decreased whereas it's voltage-dependent kinetic properties had no change. This phenomenon may be one of the mechanisms of atrial electrophysiological remodeling in chronic atrial fibrillation.

Atrial Fibrillation ; metabolism ; physiopathology ; Calcium Channels, L-Type ; physiology ; Female ; Heart Atria ; Humans ; Male ; Myocytes, Cardiac ; metabolism ; physiology ; Patch-Clamp Techniques

OBJECTIVETo evaluate the prevalence of TET2 gene mutation in acute myeloid leukemia (AML) patients, and analyze their clinical characteristics and prognosis.

METHODSPolymerase chain reaction (PCR) and direct sequencing were used to sequence exon 3 to 11 of TET2 gene.

RESULTSAmong 96 AML patients, TET2 gene mutation was detected in 13 (13.54%) patients (95%CI 6.70% - 20.38%). The median age was 54 years in mutated group and 41 years in unmutated group (P = 0.010). Mutated and unmutated patients did not significantly differ in gender, white blood cells (WBC) count at diagnosis, platelet count, PB and BM blast percentage and chromosome karyotype, excepting for hemoglobin level 84 (70 - 108) g/L in mutated group versus 70 (55 - 87) g/L in unmutated group (P = 0.032). TET2 gene mutation had no significant correlation with C-KIT, FLT3, JAK2V617F mutations, but did with NPM1 mutation. TET2 mutated patients had lower CR1 rate and 2-year overall survival than unmutated in non-M(3) patients (P < 0.05).

CONCLUSIONSTET2 gene mutation is more prevalent in older AML patients and has a certain correlation with clinical characteristics and outcome. It may be a molecular marker for poor prognosis in AML.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA Mutational Analysis ; DNA-Binding Proteins ; genetics ; Exons ; Female ; Humans ; Karyotype ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Proto-Oncogene Proteins ; genetics ; Young Adult

OBJECTIVETo investigate the mechanism by which heat shock protein 90 (HSP90) regulates 26S proteasome in hyperthermia.

METHODSHyperthermic HepG2 cell models established by exposure of the cells to 42 degrees celsius; for 3, 6, 12, and 24 h were examined for production of reactive oxygen species (ROS) and cell proliferation, and the changes in Hsp90α and 26S proteasome were analyzed.

RESULTSROS production in the cells increased significantly after hyperthermia (F=28.958, P<0.001), and the cell proliferation was suppressed progressively as the heat exposure time extended (F=621.704, P<0.001). Hyperthermia up-regulated Hsp90α but decreased the expression level (F=164.174, P<0.001) and activity (F=133.043, P<0.001) of 26S proteasome. The cells transfected with a small interfering RNA targeting Hsp90α also showed significantly decreased expression of 26S proteasome (F=180.231, P<0.001).

CONCLUSIONThe intracellular ROS production increases as the hyperthermia time extends. Heat stress and ROS together cause protein denature, leading to increased HSP90 consumption and further to HSP90 deficiency for maintaining 26S proteasome assembly and stability. The accumulation of denatured protein causes unfolded protein reaction in the cells to eventually result in cell death.

HSP90 Heat-Shock Proteins ; metabolism ; Hep G2 Cells ; Hot Temperature ; Humans ; Proteasome Endopeptidase Complex ; metabolism ; RNA, Small Interfering ; genetics ; Reactive Oxygen Species ; metabolism ; Up-Regulation

OBJECTIVETo measure the concentration of polybrominated diphenyl ethers (PBDEs) in human breast milk from a hospital in Shanghai and analyze the influencing factors.

METHODSForty-eight puerperal from a hospital in Shanghai were selected to answer a questionnaire, and their breast milk samples were collected from September 2006 to April 2007. All the puerperal were singleton pregnancies, excluding high blood pressure, diabetes, HIV infection and adverse medical history. Seven congeners (BDE-28, BDE-47, BDE-100, BDE-99, BDE-154, BDE-153 and BDE-183) were measured by gas chromatography-negative chemical ionization-mass spectrometry and the influencing factors were analyzed.

RESULTSThe median of total PBDEs concentration in breast milk was 8.81 ng/g lipid weight (lw), and the range was 1.92 - 41.55 ng/g lw. The detection rates of seven congeners (BDE-28, BDE-47, BDE-100, BDE-99, BDE-154, BDE-153, BDE-183) were 98% (47/48), 96% (46/48), 86% (41/48), 90% (43/48), 83% (40/48), 98% (47/48), 90% (43/48), and the median of them was 0.88, 0.99, 0.97, 1.39, 1.14, 2.17, 1.41 ng/g lw, respectively. LogΣ(7PBDEs) in breast milk from mothers with different education levels were divided into junior high school or lower (0.89 ± 0.24), senior high school (1.02 ± 0.17), junior college or higher (1.08 ± 0.28). LogΣ(7PBDEs) in breast milk from mothers with different income levels were divided as < 1000 yuan (0.89 ± 0.12), 1000 - 3000 yuan (1.01 ± 0.24), > 3000 yuan (1.13 ± 0.21). Correlation analysis showed that both the mothers' education level (r = 0.322, P < 0.05) and income level (r = 0.388, P < 0.05) have a positive correlation to PBDEs levels in breast milk.

CONCLUSIONIt is very common to detect PBDEs in human breast milk, however, the education and income levels of the mothers may be the influencing factors.

Adolescent ; Adult ; China ; Environmental Exposure ; Female ; Halogenated Diphenyl Ethers ; analysis ; Humans ; Milk, Human ; chemistry ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo explore the concentration of polybrominated diphenyl ethers (PBDEs) in umbilical cord serum and analyze the influence of exposure to PBDEs during fetal stage on newborn birth outcomes in Shanghai.

METHODSFifty delivery women in a Shanghai hospital were surveyed by questionnaire, and the umbilical cord serum were collected from September 2006 to April 2007. All the delivery women were singleton pregnancies, excluding high blood pressure, diabetes, HIV infection and adverse medical history. Seven congeners (BDE-28, BDE-47, BDE-99, BDE-100, BDE-153, BDE-154 and BDE-183) were measured by gas chromatography-negative chemical ionization-mass spectrometry and the influencing factors were analyzed.

RESULTSNewborns' length, weight, chest circumference, head circumference and body mass index (BMI) were (50.15 ± 0.75) cm, (3.49 ± 0.42) kg, (34.76 ± 1.51) cm, (35.03 ± 1.40) cm, (13.76 ± 1.36) kg/m(2), respectively. The median of Σ(7PBDEs) concentration in umbilical cord serum was 14.06 (1.03 - 379.73) ng/g lipid weight (lw). The detection rate of BDE-28, BDE-47, BDE-99, BDE-100, BDE-153, BDE-154 and BDE-183 were 22% (11/50), 22% (11/50), 98% (49/50), 72% (36/50), 76% (38/50), 90% (45/50) and 14% (7/50), respectively. The median (range) of PBDEs (BDE-28, BDE-47, BDE-99, BDE-100, BDE-153, BDE-154, BDE-183) congeners were < LOD (< LOD-137.20 ng/g lw), < LOD (< LOD-33.17 ng/g lw), 7.54 ng/g lw (< LOD-94.01 ng/g lw), 1.57 ng/g lw (< LOD-46.95 ng/g lw), 0.63 ng/g lw (< LOD-79.08 ng/g lw), 0.63 ng/g lw (< LOD-22.30 ng/g lw) and < LOD (< LOD-21.63 ng/g lw), respectively. The newborns' BMI showed a negative correlation with BDE-99 (r = -0.347, P < 0.05) and BDE-154 (r = -0.292, P < 0.05). BDE-99 in low-length group (≤ 50 cm, 10.59 ng/g lw) was significantly higher (t = 2.32, P = 0.03) than that in high-length group (> 50 cm, 3.60 ng/g lw).

CONCLUSIONPBDEs were commonly detected in newborns' umbilical cord serum in this study. Our findings indicated that exposure to PBDEs adversely affected the development of the newborns.

China ; Female ; Fetal Blood ; chemistry ; Humans ; Infant, Newborn ; Maternal Exposure ; Polybrominated Biphenyls ; blood ; Pregnancy ; Prenatal Exposure Delayed Effects ; Surveys and Questionnaires