OBJECTIVETo observe different effects of moxibustion on extracellular potassium ion in acupoint under physiological and pathological status and provide experimental evidence for exploring action mechanism of moxibustion on acupoint local.

METHODSForty female SD rats were randomly divided into a blank group, a blank-moxibustion group, a model group and a model-moxibustion group, 10 cases in each one. The complete Freund's adjuvant(CFA) was adopted to establish model of adjuvant arthritis (AA) in the model group and model-moxibustion group. No treatment was given in the blank group and model group while moxibustion was applied at "Zusan-li" (ST 36) for 30 min in the blank-moxibustion group and model-moxibustion group. The tissue fluid in "Zusanli" (ST 36) was collected with microdialysis and real-time analyzed by electrolytic analyzer. The change of concentration of potassium ion in "Zusanli" (ST 36) was observed.

RESULTS(1) Under physiological status, the concentration of extracellular potassium ion in the blank group was not changed within 150 min (P > 0.05); before the moxibustion, the concentration of extracellular potassium ion in the blank-moxibustion group was (1.21 +/- 0.31) mmol/L, and after treatment it was gradually increased and reached its peak at (2.38 +/- 0.42) mmol/L after 60 min (P < 0.05), then it was reduced. 150 min after the treatment, concentration of potassium ion was slightly higher than that before moxibustion as well as that in the blank group. The concentration in the blank-moxibustion group at 60 min was statistically significant compared with that in the blank group (P < 0.05). (2) Under pathological status, the concentration of extracellular potassium ion in the model group was not changed within 150 min, differences of which at each time point was not statistically significant (all P > 0.05). Before the moxibustion, the concentration of extracellular potassium ion was (1.09 +/- 0.12) mmol/L in the model-moxibustion group, and it was immediately increased to (1.96 +/- 0.18) mmol/L after moxibustion. 60 min and 90 min after the moxibustion, it still maintained a higher level, which was (1.87 +/- 0.29) mmol/L and (1.59 +/- 0.16) mmol/L respectively (both P < 0.05). The differences of each time point after moxibustion in the model-moxibustion group were statistically significant compared with those in the model group (all P < 0.05).

CONCLUSIONThe moxibustion could increase the concentration of potassium ion in rat's acupoint local under physiological status but time of effect is short; with moxibustion at "Zusanli" (ST 36) under pathological status, the concentration of local potassium ion is obviously increased and maintains for a long time.

Acupuncture Points ; Animals ; Arthritis, Experimental ; metabolism ; therapy ; Disease Models, Animal ; Female ; Humans ; Moxibustion ; Potassium ; metabolism ; Rats ; Rats, Sprague-Dawley

BACKGROUNDRetinal pigment epithelial (RPE) cells play an important role in the occurrence of choroidal neovascularization (CNV). Vascular endothelial growth factor (VEGF) as a positive regulatory growth factor is produced by the RPE in an autocrine or paracrine manner, promoting CNV development. Duplexes of 21 nt RNAs, known as short interfering RNAs (siRNAs), efficiently inhibit gene expression by RNA interference when introduced into mammalian cells. We searched for an efficient siRNA to interfere with VEGF expression in RPE cells and shed light on the treatment of CNV.

METHODSHuman primary RPE (hRPE) cells were cultured and identified. Three pairs of siRNAs were designed according to the sequence of VEGF 1-5 extrons and synthesized by T7 RNA polymerase transcription in vitro. To evaluate the inhibitory activity of T7-siRNAs, hRPE cells were transfected via siPORT Amine. The interfering effect of T7-siRNAs in hRPE cells was examined by semiquantitative reverse transcription-polymerase chain reaction and immunofluorescence.

RESULTSThree pairs of T7-siRNAs synthesized by in vitro transcription with T7 RNA polymerase suppressed VEGF gene expression with efficiency from 65% to 90%. T7-siRNA (B), targeted region at 207 nt to 228 nt and double stranded for 21 nt with 2 nt UU 3' overhangs, was the most effective sequence tested for inhibition of VEGF expression in hRPE cells. Compared with nontransfected cells, the mean fluorescence in hRPE cells transfected with T7-sRNAs was significantly less (P < 0.01). siRNA with a single-base mismatch and ssRNA(+) did not show suppressing effect. Furthermore, it was found that siRNAs had a dose dependent inhibitory effect (5 to 10 pmol).

CONCLUSIONT7-siRNA can effectively and specifically suppress VEGF expression in hRPE cells and may be a new way to treat CNV.

Base Sequence ; Cells, Cultured ; Choroidal Neovascularization ; therapy ; DNA-Directed RNA Polymerases ; metabolism ; Humans ; Molecular Sequence Data ; Pigment Epithelium of Eye ; cytology ; metabolism ; RNA Interference ; RNA, Small Interfering ; biosynthesis ; pharmacology ; Transcription, Genetic ; Vascular Endothelial Growth Factor A ; antagonists & inhibitors ; genetics ; Viral Proteins ; metabolism

OBJECTIVETo investigate the distribution characteristics and influencing factors of mental workload of teachers in primary schools.

METHODSNational Aeronautics and Space Administration-Task Load Index (NASA-TLX) was used to assess the mental workload levels for 397 teachers of primary schools in a city.

RESULTSThe mental workload (64.34+10.56) of female teachers was significantly higher than that (61.73+ 9.77) of male teachers (P<0.05). The mental workload (65.66+10.42) of "-35" years old group was the highest. When age of teachers was younger than 35 years old, there was a positive correlation between the mental workload and age (r=0.146, P<0.05). When age of teachers was older than 35 years old, there was a negative correlation between the mental workload and age (r=-0.190, P<0.05). The teachers with higher education level felt higher mental workload (unstandardized coefficients B=1.524, standardized coefficients /=0.111, P<0.05). There was a positive correlation between the mental workload and working hours per day (unstandardized coefficients B =4.659, standardized coefficients/3 =0.223, P<0.001).

CONCLUSIONMental workload of the teachers in primary schools is closely related to age, educational level and work hours per day. Work hours per day is an important risk factor for mental workload. Reducing work hours per day (8 hours) is an effective measure of alleviating the mental workload of teachers in primary schools.

Adolescent ; Adult ; Aged ; Faculty ; Female ; Humans ; Male ; Middle Aged ; Schools ; Stress, Psychological ; epidemiology ; Surveys and Questionnaires ; Workload ; psychology ; Young Adult

AIMTo demonstrate the specific killing of folate receptor (FR)-positive tumor cells can be achieved by folate-targeted penicillin-G amidase (PGA) combined with its prodrug substrate N-(phenylacetyl) doxorubicin (DOXP).

METHODSFolic acid was covalently linked to PGA and folate content value was determined by quantitative UV spectrophotometry. The ability of folate conjugated PGA to hydrolyze DOXP was measured by RP-HPLC. Visual demonstration of uptake by FR (+) HeLa and SKOV3 cells was detected by using FITC labeled folate-PGA and a fluorescence microscopy. The cytotoxicity of DOXP towards the cells in the presence or absence of folate-PGA was assayed by using MTT method.

RESULTSThe folate-PGA has a specific activity of 29. 8 U x mg(-1) (protein). FR selectivity was confirmed by fluorescence microscopy. The combination of DOXP prodrug with folate-PGA generated higher cytotoxicity towards the FR (+) cells than free doxorubicin. The IC50 was 0.72 micromol x L(-1) for HeLa cells and 0.75 micromol x L(-1) for SKOV3 cells, respectively. Further, the enhanced cytotoxicity reduced greatly with the addition of free folic acid.

CONCLUSIONFolate conjugated PGA did not significantly compromise PGA catalytic activity and enabled binding prodrug-activating enzyme PGA to folate receptor expressing cells, and increased the sensitivity of the cells to doxorubicin followed by administration of its prodrug substrate.

Antibiotics, Antineoplastic ; pharmacology ; Carrier Proteins ; metabolism ; Cell Line, Tumor ; Doxorubicin ; analogs & derivatives ; pharmacology ; Drug Delivery Systems ; Female ; Folate Receptors, GPI-Anchored ; Folic Acid ; chemistry ; pharmacology ; HeLa Cells ; Humans ; Inhibitory Concentration 50 ; Ovarian Neoplasms ; pathology ; Penicillin Amidase ; chemistry ; pharmacology ; Prodrugs ; pharmacology ; Receptors, Cell Surface ; metabolism

Objective:To investigate the relationship between the expression of EZH2 and the prognosis of elderly patients with rectal cancer and the mechanism of EZH2 in the progression of rectal cancer.Methods:36 cases of elderly patients with rectal cancer who underwent surgery in our hospital from January to June 2010 were included in this study.The expression of EZH2 in colorectal cancer tissues and adjacent tissues was analyzed and the differences were analyzed.All the patients were divided into positive and negative groups according to the expression level of EZH2.The correlation factors and risk factors of EZH2 expression were analyzed.All patients were followed up for 1 year to assess the quality of life of patients with prognosis and to analyze the correlation with the expression of EZH2;5-year follow-up of patients in both groups compared to 5-year progression-free survival rate of progression-free survival And overall survival(PFS) and overall survival (OS) survival curves were recorded.Results:The positive rate of EZH2 was 69.44％ in CRC and 38.89％ in cancerous tissues,and the positive rate of EZH2 in cancer tissues was significantly higher than that in adjacent tissues (P＜0.01).Age,pathologic type,The positive rate of EZH2 expression in lymph node metastasis group was significantly higher than that in lymph node metastasis group (P＜0.05).Logistic regression analysis showed that EZH2 expression was an independent risk factor of CRC lymph node metastasis.The QLQ-30 quality of life score was significantly higher in EZH2 low-expression group than that in EZH2 high-expression group (P＜0.01).The expression level of EZH2 was negatively correlated with QLQ-30 quality of life score (r=-0.573,P＜0.01).The 5-year progression-free survival rate of the 5-year EZH2 low-expression group was 86.67％ and the 5-year overall survival rate was 93.33％.The 5-year progression-free survival rate was 52.38％ 61.90％.The 5-year progression-free survival rate and 5-year overall survival rate of EZH2 low expression group were significantly higher than those of EZH2 high expression group (P＜0.05).Conclusion:The high expression of EZH2 is closely related to the progression of rectal cancer.It can be used as an important predictor of prognosis quality of life,progression-free survival time and overall survival time.

Objective:To study the expression of tumor suppressor factors SEMA3F,CYLD and miR-454 in rectal cancer.Methods:21 patients with colorectal cancer admitted in our hospital from January to March in 2016 were selected as the study materials.The tumor inhibitoryfactor SEMA3F in rectal cancer tissue,cancerous surrounding tissue and normal rectum tissue were detected respectively,CYLD and miR-454,and to analyze the correlation between tumor suppressor factors SEMA3F,CYLD and miR-454,and to explore their roles in the carcinogenesis and progression of rectal cancer.Results:The expression of SEMA3F,CYLD and miR-454 in cancer tissue,paracancer tissue and normal tissue were statistically significant (P＜0.01).The levels of SEMA3F and CYLD were significantly lower in patients with metastasis than those without metastasis,and miR-454 was significantly higher than that in patients without metastasis (P＜0.01).The expression level of SEMA3F and CYLD in CRC tissues was significantly higher than that in moderately and poorly differentiated tissues,and the expression of miR-454 was significantly lower than that in moderately and poorly differentiated ones (P＜0.01).There was significant negative correlation between tumor suppressor CYLD and miR-454 (r=-0.971,P＜0.01).There was a significant negative correlation between tumor inhibitory factor SEMA3F and miR-454(r=-0.955,P＜0.01).Conclusion:The tumor suppressor factors SEMA3F and CYLD play an important role in inhibiting the formation and proliferation of cancer cells in colorectal cancer.The expression of tumor suppressor factors SEMA3F and CYLD in colorectal cancer is high in the early stage and gradually declines with the progression of the disease.MiR-454 can promote the growth of rectal cancer cells The expression of tumor suppressor factors SEMA3F,CYLD and miR-454 in colorectal carcinoma were significantly higher than those in SEMA3F and CYLD.The expression of tumor suppressor factors such as SEMA3F,CYLD and miR-454 were significantly correlated with the progression and prognosis of rectal cancer The evaluation has important reference value.

Ornithine decarboxylase (ODC) is the rate-limiting enzyme in polyamine biosynthesis and a target for chemoprevention. Hydroxydibenzoylmethane (HDB), a derivative of dibenzoylmethane of licorice, is a promising chemopreventive agent. In this paper, we investigated whether HDB would inhibit the ODC pathway to enhance apoptosis in human promyelocytic leukemia HL-60 cells. We found ODC enzyme activity was reduced during HDB treatment. Overexpression of ODC in HL-60 parental cells could reduce HDB-induced apoptosis, which leads to loss of mitochondrial membrane potential (Deltapsim), through lessening intracellular ROS. Furthermore, ODC overexpression protected cytochrome c release and the activation of caspase-3 following HDB treatment. The results demonstrated HDB-induced apoptosis was through a mechanism of down-regulation of ODC and occurred along a ROS-dependent mitochondria-mediated pathway.
Apoptosis/*drug effects ; Caspase 3/metabolism ; Chalcones/metabolism/*pharmacology ; Chemoprevention ; Cytochromes c/biosynthesis/secretion ; Down-Regulation ; Gene Expression ; HL-60 Cells ; Humans ; Immunoblotting ; Leukemia, Myeloid/*enzymology/pathology ; Membrane Potential, Mitochondrial/drug effects ; Mitochondria/enzymology ; Ornithine Decarboxylase/antagonists & inhibitors/genetics/*metabolism ; Reactive Oxygen Species/analysis/metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the prevalence of pulmonary surfactant associated pathway genes functional variants in Chinese population.

METHODUsing a cohort of 258 mixed ethnic population of Han and Zhuang, we pooled DNA samples from 146 term male infants and 112 term female infants and then used an Ill umina next generation sequencing platform to perform the complete exonic resequencing in 6 target genes:surfactant protein-B (SFTPB), surfactant protein-C (SFTPC), ATP-binding cassette transporter A3 (ABCA3), lysophospholipid acyltransferase 1 (LPCAT1), choline phosphotransferase 1 (CHPT1), phosphate cytidylyltransferase 1, choline, beta (PCYT1B). Collapsing methods was used to determine the functional allele frequency.

RESULT(1) Altogether, 128 variants were found, including 44 synonymous variants, 66 nonsynonymous variants and 18 insertions-deletions. Of these, 28 variants were predicted to alter protein function. Two of these variants were seen twice, the rest variants were only seen once, for a total of 30 functional alleles; (2) ABCA3 had the most functional variants in both male and female groups with the minor allele frequencies of 0.014 (1.4%) and 0.04 (4%), respectively. The total functional allele frequencies of 6 genes were 0.041 (4.1%) and 0.08 (8%) in the two groups, respectively (P = 0.06).

CONCLUSION(1) Functional variants in pulmonary surfactant associated pathway genes are present in the mixed Han-Zhuang population. (2) ABCA3 contained the most functional variants suggesting that ABCA3 could contribute significantly to neonatal respiratory distress syndrome and other lung disease.

1-Acylglycerophosphocholine O-Acyltransferase ; genetics ; metabolism ; ATP-Binding Cassette Transporters ; genetics ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Female ; Gene Frequency ; Genetic Association Studies ; Genetic Predisposition to Disease ; Genetic Variation ; Genotype ; Humans ; Infant, Newborn ; Male ; Pulmonary Surfactant-Associated Protein C ; genetics ; Pulmonary Surfactant-Associated Proteins ; genetics ; Respiratory Distress Syndrome, Newborn ; ethnology ; genetics

OBJECTIVETo explore the changes and the clinical significance of 5-hydroxytryptamine (5-HT), dopamine (DA) levels in serum and cerebrospinal fluid (CSF) of patients with delayed encephalopathy (DEACMP) after acute carbon monoxide poisoning.

METHODSThe dynamic detection of 5-HT and DA levels in serum and CSF from 42 patients with DEACMP was performed with high performance liquid chromatography (HPLC). The condition changes of patients with DEACMP were analyzed with three types of scales: the activity of daily living scale (ADL), information memory concentration test (IMCT) and Hasegawa's dementia scale (HDS); these changes were compared with those from 38 other encephalopathy patients and 38 non-encephalopathy patients, respectively.

RESULTSBefore treatment, the serum 5-HT and DA levels [(662.61 ± 178.50) and (155.74 ± 60.32) nmol/L, respectively] of DEACMP group were both significantly lower than those [(914.08 ± 198.04) and (225.70 ± 48.53) nmol/L] of non-encephalopathy group (P < 0.05); the serum DA level of DEACMP group was also significantly lower than that [(243.57 ± 66.94) nmol/L] of other encephalopathy group (P < 0.05); the serum 5-HT level of DEACMP group was not significantly different from that [(729.54 ± 299.87) nmol/L] of other encephalopathy group (P > 0.05). After treatment, the serum 5-HT and DA levels [(714.08 ± 170.47) and (192.18 ± 33.07 nmol/L, respectively)] of DEACMP group elevated to various extent, but only serum DA level was significantly higher than that before treatment (P < 0.05). Before treatment, the CSF 5-HT and DA levels of DEACMP group were significantly lower than those of non-encephalopathy group and those of other encephalopathy group (P < 0.05). After treatment, the CSF 5-HT level (232.44 ± 54.28 nmol/L) was similar to normal level and significantly higher than that before treatment (P < 0.05); the CSF DA level [(56.83 ± 12.85) nmol/L] of DEACMP group increased only slightly (P > 0.05). In DEACMP group, ADL score (50.64 ± 7.23), HDS score (8.55 ± 8.08) and IMCT score (4.95 ± 7.30) before treatment were significantly different from those (8.5 ± 8.08, 4.95 ± 7.30 and 15.64 ± 10.90) after treatment (P < 0.01). In DEACMP group, there wasa negative correlation between DA level changes and HDS score changes, when the DA levels and HDS scores before treatment were compared with those after treatment (P < 0.05).

CONCLUSIONThe dynamic changes of 5-HT and DA levels in serum and CSF of patients with DEACMP consisted basically with the patient's condition change. The dynamically detected 5-HT and DA levels can be used as the biological indicators to reflect the condition change and treatment effects of DEACMP patients.

Adult ; Aged ; Aged, 80 and over ; Brain Diseases ; blood ; cerebrospinal fluid ; etiology ; Carbon Monoxide Poisoning ; blood ; cerebrospinal fluid ; complications ; Case-Control Studies ; Dopamine ; blood ; cerebrospinal fluid ; Female ; Humans ; Male ; Middle Aged ; Neurotoxicity Syndromes ; blood ; cerebrospinal fluid ; etiology ; Serotonin ; blood ; cerebrospinal fluid

OBJECTIVETo study the feasibility of endolymphatic visualization and the diagnosis of Meniere's disease by applying intratympanic gadolinium administration through the tympanic membrance and three-dimensional fluid-attenuated inversion recovery magnetic resonance imaging (3D-FLAIR MRI). To study the relationship between the endolymphatic hydrops visualized by MRI and audio-vestibular functional tests, such as pure tone audiometry (PTA), electrocochleography (EcoG), caloric test and vestibular evoked myogenic potential (VEMP).

METHODSWith a three Tesla magnetic resonance imaging (MRI) unit, 3D-FLAIR imaging was performed 24 hours after intratympanic gadolinium through the tympanic membrance in 32 patients with clinically diagnosed unilateral Meniere's Disease. We visualized the enhanced imaging of perilymphatic space in bilateral cochlea, vestibular and (or) canal, scoring scala tympani and scala vestibule of bilateral cochlear basal turn respectively and measuring the developing area of bilateral vestibule and the signal intensity ratio (SIR) between the vestibule and the brain stem subjectively. PTA, EcoG, caloric test and VEMP were performed. The relationship between the endolymphatic hydrops visualized by MRI and audio-vestibular functional tests were studied.

RESULTSThe gadolinium appeared in almost all parts of the perilymph in cochlea, vestibular and (or) canals in all 32 patients' inner ears, so the endolymphatic space was clearly shown on 3D-FLAIR imaging. The scala vestibuli score value between the affected side and the healthy side were statistically significant (Z = 4.309, P < 0.05) . The developing vestibular area between the affected side and the healthy side [(6.04 ± 2.89) mm(2), (8.28 ± 3.04)mm(2)] were statistically significant (t = 3.322, P < 0.05) . Abnormal vestibular evoked myogenic potentials were significantly correlated with the developing vestibular area of the affected side (F = 11.96, P < 0.05) . Abnormal electrocochleography were significantly correlated with scala vestibuli score value of cochlear basal turn in the affected side (Z = 3.17, P < 0.05) . No significant correlation was found between the scala vestibuli score value or the developing vestibular area and caloric test or PTA findings.

CONCLUSIONS3D-FLAIR MRI with intratympanic gadolinium injection through the tympanic membrance can discriminate the border between the perilymph and the endolymph and show endolymphatic hydrops. This method may provide radiographic reference for the diagnosis of Meniere's disease. The results of VEMP and electrocochleography might have appropriate correlation with degree of vestibular and cochlear hydrops.

Audiometry, Evoked Response ; Audiometry, Pure-Tone ; Caloric Tests ; Cochlea ; Contrast Media ; Ear, Inner ; Endolymph ; Endolymphatic Hydrops ; complications ; diagnosis ; Gadolinium DTPA ; Humans ; Imaging, Three-Dimensional ; Injections ; Magnetic Resonance Imaging ; Meniere Disease ; complications ; Perilymph ; Vestibular Evoked Myogenic Potentials ; Vestibule, Labyrinth