Objective To study the effects of hydrogen dioxide (oxygen free radical donator) and vitamin C (oxygen free radical scavenger) on the electric current of large conductance calcium -activated potassium channels (BKCa channels) in isolated outer hair cells in aging guinea pigs .Methods Acute enzyme was used to isolat outer hair cells of aging guinea pigs ,in which of BKCa channel's electric current was observed and recorded by whole-cell recording mode of patch -clamp .After recording the stable and normal electric current of BKCa channels ,added H2 O2 dilution (0 .2 mmol/L) 40 μl in the 2 ml chambers within freshly isolated outer hair cells so that the concen‐tration of H2 O2 in the balneum would be 4 μmol /L .The groups(n=5) received individually vitamin C solution (5 mg/ml) 0 ,10 ,20 ,40μl in the 2 ml chambers within freshly isolated outer hair cells so that the concentration of vi‐tamin C in the balneum would be 0 ,25 ,50 ,100 μg /ml ,observing and recording the effects of different concentration of vitamin C to electric current of BKCa channels .Results ①In the w hole-cell mode of patch -clamp ,the rapid activation and non-deactivation electric current with a string of large amplitude was recorded ,above -40~ -30 mV activation voltage .The electric current increased with the increasing membrane potential .The amplitude in‐creased continuously and performed characteristics of outward rectification .When the concentration of IbTX was 100 nmol /L ,the activity of the channel was completely blocked and confirmed BKCa channel's electric current .②Medication within three minutes ,when VT was +50 mV ,the BKCa channels'the maximum peak current densities of 4 μmol /L H2 O2 group rose from 22 .09 ± 0 .27 PA /PF to 43 .53 ± 1 .09 PA/PF ,amplification was 97 .06% .In H2 O2 4 μmol /L + vitamin C with different concentrations as 25 ,50 ,100 μg/ml groups ,the BKCa channels'elec‐tric current performed about concentration-dependent inhibition ,and electric current's amplitude and peak current density decreased with the increasing concentration of vitamin C ,the I-V curves were reduced .However ,this still could not be recovered to the normal levels .Conclusion The oxygen free radical /BKCa exists in the process .The vitamin C as oxygen free radical scavenger can reverse the process to a large extent .

Objective The purpose of this study was to investigate the correlation of high sensitivity C-reactive protein (hsCRP) and fibrinogen with carotid artery arteriosclerosis of patients with cerebral infarction. Methods One hundred and thirteen patients with cerebral infarction were assigned as study group, and 102 healthy persons as control group. The levels of serum hsCRP and Fib in the two groups were measured. The carotid artery arteriosclerosis and carotid intimal-medial thickness (IMT) were examined by color Doppler and B-ultrasound. Results The value of IMT between study group and control group was statistically significant. The positive rates of carotid artery arteriosclerosis plaque and vulnerable plaque in study group were significantly higher than those in control group (all<0.05) . The level of serum hsCRP was significantly higher in study group than that of control ( <0.05) . The level of serum Fib between study group and control group was not statistically significant ( >0.05) . Conclusion The level of hsCRP was closely related to the degree of carotid artery arteriosclerosis and the occurrence and development of cerebral infarction. But the level of Fib was not closely related to the degree of carotid artery arteriosclerosis.

Objective To evaluate the application of prevention technique for mother‐to‐child transmission of human immunode‐ficiency virus(HIV) in Laibin City .Methods Prevention techniques for mother‐to‐child transmission of HIV were applicated in the city .Women in pregnancy test received acquired immunodeficiency syndrome(AIDS) counseling and test at the same time .Measures were taken to prevent the mother‐to‐child transmission of HIV for women who confirmed HIV positive ,including the use of antiret‐roviral drugs ,safety midwifery and feeding guidance ,and so on .Results The average HIV counseling and testing rate was 99 .1%(176 001/177 738) for the past 5 years .269 cases were HIV positive ,and the positive rate was 0 .15% (269/176 001) .Via health advisory guidance ,pregnant women diagnosed with AIDS understood the dangers of AIDS and the mode of its transmission ,and then free to chose weather to have the pregnancy terminated .Finally ,there were 67 pregnant women chose to terminate the preg‐nancy ,and 202 women decided to have children who were included in the prevention management system of mother‐to‐child trans‐mission .202 infants were born alive ,192 of which took full amount of antiretroviral drugs .The medication rate was 95 .05% (192/202) .42 d and 3 months after birth ,early diagnosis of HIV/AIDS was performed for the infants ,the positive rate of HIV was 0 .50% (1/202) .There were 118 infants at least 18‐months‐old who had underwent HIV test and no positive case was detected . Conclusion AIDS counseling tests in pregnancy test women is necessary ,which could detect HIV positive women .Comprehensive measures are effective methods for the prevention of mother‐to‐child transmission .

Objective:To evaluate the effect of repeated intranasal insulin on postoperative delirium (POD) in elderly patients undergoing general anesthesia.Methods:Seventy elderly patients, aged ≥65 yr, with body mass index ≤28 kg/m 2, of American Society of Anesthesiologists physical statusⅠ-Ⅲ, undergoing elective radical gastrectomy for gastric cancer under general anesthesia, were divided into 2 groups ( n=35 each) according to the random number table method: control group (group C) and insulin group (group I). In group C, normal saline 0.5 ml was administered intranasally twice a day from 2 days before surgery until the day of surgery.In group I, insulin 20 U (0.5 ml) was administered intranasally twice a day from 2 days before surgery until the day of surgery.The regional tissue oxygen saturation (rSO 2) was measured after entering the operating room (T 0), at intubation (T 1), at 1, 2 and 3 h after the start of operation (T 2-4), at the end of surgery (T 5) and at extubation (T 6). The insulin allergic reactions, nasal irritation and hypoglycemic reactions were recorded after intranasal administration of insulin or normal saline within 2 days before operation.The blood glucose concentrations were measured at T 0, T 3 and T 5.The occurrence of POD within 3 days after operation was recorded. Results:Compared with group C, the incidence of POD was significantly decreased within 3 days after operation, and the rSO 2 was increased at T 1-6 in group I ( P<0.05). The rSO 2 was significantly higher at T 1-6 than at T 0 in two groups ( P<0.05). There were no significant changes in blood glucose concentrations at T 0, T 3 and T 5 between the two groups ( P>0.05). No insulin allergic reactions, nasal irritation and hypoglycemic reactions occurred in two groups ( P>0.05). Conclusion:Repeated intranasal administration of insulin can increase the rSO 2 during operation and decrease the occurrence of POD in elderly patients undergoing radical gastrectomy for gastric cancer.

Objective To investigate the association of promoter hypermethylation of secreted frizzled-related proteins (SFRPs) in patients with colorectal cancer. Methods The promoter hypermethylation of SFRPs in 20 sporadic colorectal cancer tissues and adjacent mucosa were detected by methylation-specific PCR. The amplified DNA was subcloned into the T-A cloning vector and sequenced. Two colorectal cancer cell lines (HCT116 and SW480) were treated with 5-aza-2' deoxycytidine for demethylation. The promoter hypermethylation and protein expression of SFRPs in colorectal cancer cell lines were detected by methylation-specific PCR and Western blotting. Results It was demonstrated that the hypermethylation of SFRP 1, 2, 4 or 5 was 19/20,17/20,3/20 or 13/20in cancer tissues, respectively, whereas it was 12/20, 8/12, 1/20 or 7/20 in adjacent mucosa,respectively. SFRP 1, 2 or 5 methylation was more frequently found in cancer tissue than in adjacent mucosa (P～0.05). Methylation of SFRP 1, 2, 4 and 5 were found in HCT116 cell line, but only SFRP1 and SFRP2 were found in SW480 cell line. There was a negative correlation between protein expression and methylation of SFRPs. The Western blotting revealed that SFRP protein re-expressedafter it treated with 5-aza-2' deoxyeytidine. Conclusion Methylation of SFRP 1, 2 or 5 gene is associated with the evolution of eolorectal cancer, and is closely related to silencing expression.

Objective To evaluate the effectiveness and safety of multimodal postoperative analgesia of thoracotomy for malignant tumor excision.Method Sixty patients undergoing thoracotomy for malignant tumor excision were recuited.They were randomized into four groups:group A receiving single flurbiprofen axetil injection,group B using electronic analgesia pump for sufentanil PCIA,group C using electronic analgesia pump for dezocine PCIA and group D receiving sufentanil controlled by PCA-TCI system.Vital signs,artery lood gas analysis,PrinceHenry score,Ramsay score,remedial measure and untoward reactions were recorded.Results The falling down degree of Prince-Henry score of Group D was higher than those of Groups A,B and C (P ＜ 0.05).Reduction of Ramsay score of Group D was higher than those of Groups A,B and C (P ＜ 0.05).The number of the patients in Group D not undergoing Bolus remedial measure was larger than that of those in Groups B and C (P ＜ 0.05).Conclusion When referring to the effectiveness and safety of the models administrated the thoracotomy for malignant tumors excision,the analgesia model of PC A-TCI sufentanil (plasma target concentration:0.12～ 0.14 ng/ mL) used in Group D is superior to other models used in Groups A,B and C.Moreover,individual analgesia can be reached by the PCA-TCI sufentanil model.

Objective To observe the effect of Qi-strengthening and blood-activating Chinese patent medicine Qigui Ershen Granules on the carotid intima-media thickness(IMT ) , atheromatous plaque scores, serum fibroblast growth factor 23 (FGF23) and Klotho protein levels, and oxidation- and inflammation-associated indicators in carotid atherosclerosis patients. Methods Fifty-two carotid atherosclerosis patients were randomized into Chinese medicine group and western medicine group, 26 cases in each group. Chinese medicine group was treated with Qigui Ershen Granules orally, and western medicine group was treated with Atorvastatin Calcium Tablets orally. The mediation for the two groups lasted for 24 continuous weeks. Carotid ultrasonography was performed before and after treatment for the examination of carotid IMT and plaque Crouse scores. Double antibody sandwich enzyme-linked immunosorbent assay(ELISA) was applied for the detection of serum Klotho, FGF23, interleukin 1(IL-1) and tumor necrosis factorα(TNF-α) levels, and radio-immuno-precitation method was used for the assay of serum reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) levels. The clinical efficacy of the two groups was evaluated by the scores of Qi deficiency syndrome and blood stasis syndrome before and after treatment. Results (1) In western medicine group, 5 cases dropped out and were excluded, and a total of 21 cases completed the trial; in Chinese medicine group, 3 cases dropped out and were excluded, and a total of 23 cases completed the trial.(2) After treatment for 24 continuous weeks, IMT and Crouse scores of the plaque in the two groups were obviously reduced(P < 0.01 compared with those before treatment) , but the differences of IMT and the scores between the two groups were insignificant after treatment(P > 0.05). (3) Serum Klotho protein level was increased while FGF23 was decreased in Chinese medicine group after treatment (P < 0.01 compared with those before treatment); no obvious changes of serum Klotho protein and FGF23 levels were found in western medicine group before and after treatment(P > 0.05). The effects of Chinese medicine on increasing Klotho protein level and decreasing FGF23 level were superior to those of western medicine (P<0.01). (4) After treatment, serum IL-1, TNF-α, ROS and MDA levels were decreased and serum SOD level was increased in the two groups (P < 0.01 compared with those before treatment). The differences of the above indexes were insignificant between the two groups after treatment(P > 0.05).(5) The scores of Qi deficiency syndrome and blood stasis syndrome in Chinese medicine were decreased after treatment (P < 0.01), but showed no significant changes in western medicine group (P > 0.05). Chinese medicine group had better effect on improving the scores of Qi deficiency syndrome and blood stasis syndrome than western medicine group(P < 0.01).(6) After treatment, the total effective rate for improving Qi deficiency syndrome and blood stasis syndrome in Chinese medicine group was 82.61%, 78.26%, and that in western medicine group was 28.57%, 14.28%respectively, the difference being significant (P<0.01). Conclusion Qi-strengthening and blood-activating Qigui Ershen Granules have certain effects on counteracting atherosclerosis, inflammatory aging and oxidation.

To investigate the role of volatile components in the compound and to find the substance foundation of Gualou Guizhi decoction (GLGZD) for curing extremities spasticity after stroke. The chemical compositions of essential oil, obtained by hydrodistillation from Gualou Guizhi decoction and its major constituting herbs (Trichosanthis Radix, Paeoniae Alba Radix, Cinnamomi Ramulus, Zingiberis Recens Rhizoma, Glycyrrhizae Radix, Ziziphi Jujubae Fructus) were analyzed by GC-MS to evaluate the correlativity between volatile components of GLGZD and its major constituting herbs, and volatile components after oral administration of GLGZD in the rats' brain. Volatile components of GLGZD are mainly derived from Cinnamomi Ramulus, Zingiberis Recens Rhizoma, Ziziphi Jujubae Fructus, Trichosanthis Radix. The volatile components in the brain is mostly derived from radix trichosanthis. Compared with individual herbs of GLGZD, the dissolution of the components increase or new components appear after compatibility of six herbs. Adminstrated with GLGZD, the results point out that volatile components in the brain play a neuroprotective role through passing the brain.
Animals ; Brain ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gas Chromatography-Mass Spectrometry ; Male ; Rats ; Rats, Sprague-Dawley ; Volatile Organic Compounds ; chemistry ; pharmacology

A qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS) was developed for identification of multi-constituents and an analytical method was developed for simultaneously determining 4 major compounds (rutin, isoquercitrin, kaempferol-3-0-rutinoside, and astragalin) in Tetrastigma hemsleyanum Diels et Gilg. The HPLC-Q-TOF-MS assay was performed on a Welch Ultimate XB-C18 column (4.6 mm x 150 mm, 5 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% Formic acid (B) in gradient mode at a flow rate of 0.8 mL x min(-1). The column temperature was at 30 degrees C, and negative ion mode was used for TOF-MS. The UPLC-QqQ-MS assay was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% formic acid (B) in gradient mode at a flow rate of 0.25 mL x min(-1). The column temperature was at 45 degrees C, and MRM mode was used for QqQ-MS. Based on the retention time and MS spectra, 24 compounds were identified or tentatively characterized by comparing with reference substances or literatures. For quantitative the linear range of 4 detected compounds were good (r > 0.9966), and the overall recoveries ranged from 98.27% to 101.58%, with the RSD ranging from 3.15% to 5.88%. The results indicated that new approach conbined HPLC-Q-TOF-MS and UPLC-QqQ-MS was applicable in qualitative and quantitative quality control of Tetrastigma hemsleyanum.
Acetonitriles ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Formates ; chemistry ; Tandem Mass Spectrometry ; methods ; Vitaceae ; chemistry ; Water ; chemistry

OBJECTIVETo clone SABATH methyltransferase (rLjSABATHMT) gene in Lonicera japonica var. chinensis, and compare the gene expression and intron sequence of SABATH methyltransferase orthologous in L. japonica with L. japonica var. chinensis. It provide a basis for gene regulate the formation of L. japonica floral scents.

METHODThe cDNA and genome sequences of LjSABATHMT from L. japonica var. chinensis were cloned according to the gene fragments in cDNA library. The LjSABATHMT protein was characterized by bioinformatics analysis. SABATH family phylogenetic tree were built by MEGA 5.0. The transcripted level of SABATHMT orthologous were analyzed in different organs and different flower periods of L. japonica and L. japonica var. chinensis using RT-PCR analysis. Intron sequences of SABATHMT orthologous were also analyzied.

RESULTThe cDNA of LjSABATHMT was 1 251 bp, had a complete coding frame with 365 amino acids. The protein had the conservative SABATHMT domain, and phylogenetic tree showed that it may be a salicylic acid/benzoic acid methyltransferase. Higher expression of SABATH methyltransferase orthologous was found in flower. The intron sequence of L. japonica and L. japonica var. chinensis had rich polymorphism, and two SNP are unique genotype of L. japonica var. chinensis. The motif elements in two orthologous genes were significant differences.

CONCLUSIONThe intron difference of SABATH methyltransferase orthologous could be inducing to difference of gene expression between L. japonica and L. japonica var. chinensis. These results will provide important base on regulating active compounds of L. japonica.

Base Sequence ; Cloning, Molecular ; Computational Biology ; Evolution, Molecular ; Gene Expression Regulation, Plant ; Lonicera ; enzymology ; genetics ; Methyltransferases ; genetics ; Nucleotide Motifs ; Phylogeny ; Sequence Alignment ; Sequence Analysis