To provide accurate information on geographic distribution of crude drug Sailonggu in the plateau, we identified zokor species (Eospalax spp.) in Qinghai-Tibet Plateau using molecular methods. Based on the mitochondrial cytochrome B (cytb) gene sequences, we then extracted haplotypes from these sequences and reconstructed phylogenetic trees for the haplotypes using both maximum likelihood (ML) and Bayesian inference (BI) methods. Based on the trees, the species of each sample were determined. Five hundred and three samples from 35 populations were sequenced and their whole cytb sequences (1140 bp) were obtained. From these sequences 150 haplotypes were detected, in which, 126 were Eospalax baileyi, 20 were E. cansus, and 4 were E. smithi of the 35 populations, 28 were E. baileyi type, 5 were E. cansus type, and the remaining 2 were mixed of E. baileyi + E. cansus (DT2) and E. baileyi + E. smithi (ZN3). The results showed that, the regions around the Qinghai lake and near the upper stream of Yellow River started at Guide could be viewed as the producing area of authentic Sailonggu, and also, the cytb gene is a powerful molecular marker to determine the species of zokors as well as for the authentication of geographic distribution of Sailonggu.
Animals ; Bone and Bones ; metabolism ; Haplotypes ; Medicine, Tibetan Traditional ; Phylogeny ; Rodentia ; classification ; genetics

Adult ; Antidotes ; administration & dosage ; therapeutic use ; Female ; Humans ; Insecticides ; poisoning ; Parathion ; poisoning ; Poisoning ; drug therapy ; Pralidoxime Compounds ; administration & dosage ; therapeutic use ; Treatment Outcome

OBJECTIVESTo construct the expression vector pGenesil-3-shRNA that can express the short hairpin RNAs (shRNA) silencing Kir6.2 gene and to study its influence on the proliferation and invasion of HepG2 cells.

METHODSTwo shRNA silencing Kir6.2 genes were transcript synthesized intracellularly by expressed templates of plasmid vector pSilence-3, and the target sequence of Kir6.2 gene was inserted into the upstream of the reporter gene in order to construct the recombinant plasmid vector pGenesil-3. Plasmids containing 2 different sequences of human Kir6.2 mRNA coding region were constructed and transfected into HepG2 cells by using lipofectamine 2000 methods. The experiment was divided into 4 groups: SK (normal), SK-HK (negative control), SK-K1 (transfected with the interfering sequence 1) and SK-K2 (transfected with the interfering sequence 2) groups. A selected single clone was cultured after screening by G418. The expression of Kir6.2 protein was detected by Western blot. MTT assay and Transwell system were used to observe the proliferation and invasion of HepG2 cells.

RESULTSThe recombinant expression plasmid pGenesil-3 was successfully constructed and underexpression of Kir6.2 gene in HepG2 cells was detected by Western blot. Underexpression of Kir6.2 gene significantly decreased the proliferation and invasion of the HepG2 cells.

CONCLUSIONshRNA can inhibit the expression of Kir6.2 gene in the HepG2 cells, and underexpression of Kir6.2 gene decreased the proliferation and invasion of the HepG2 cells.

Cell Proliferation ; Genetic Vectors ; Hep G2 Cells ; Humans ; Neoplasm Invasiveness ; Plasmids ; Potassium Channels, Inwardly Rectifying ; genetics ; RNA, Small Interfering ; genetics

To clone the 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase (TwMCT) full length cDNA from Tripterygium wilfordii, the specific primers were designed according to the transcriptome data and the LCPCR were carried out. After a series of bioinformatics analysis on the TwMCT, the MeJA induced expression content were investigated by real-time fluorescence quantification polymerase chain reaction (RT-qPCR). The result showed that the full of TwMCTcDNA was 1 318 bp nucleotides encoding 311 amino acids. The molecular weight of the deduced TwMCT protein was about 34.14 kDa and the theoretical isoelectric point was 8.65. Result of the RT-qPCR analysis indicated that the content of TwMCT mRNA expression in T. wilfordii suspension cell was rising after treating with MeJA and reached the maximum in 24 h. Cloning and analyzing TwMCT gene from T. wilfordii provided gene element for studying the function and expression regulation of secondary metabolites.
Amino Acid Sequence ; Cloning, Molecular ; Erythritol ; analogs & derivatives ; metabolism ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Nucleotidyltransferases ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary ; Sequence Alignment ; Sugar Phosphates ; metabolism ; Tripterygium ; chemistry ; enzymology ; genetics

AIM:To investigate the value of 3D-optical coherence tomography (OCT) in the diagnosis of macular disease before phacoemulsification.METHODS:Clinical records of 423 cataract patients (512 eyes) who underwent phacoemulsification combined with intraocular lens implantation in our hospital from June to December in 2015 were retrospectively analyzed.In addition to preoperative routine examination of fundus, Topcon 3D-OCT 2000 was used to examine the macula, the detection rate of macular disease was compared, risk factors of cataract combined with macular disease were analyzed.RESULTS:The OCT image results of 305 cases (384 eyes, 72.1%) were successfully obtained, 133 cases showed macular disease (146 eyes), the detection rate was 28.5% (95%CI:27.64%-29.40%);the macular disease of 35 cases (37 eyes) were detected by routine examination of fundus before operation, the detection rate was 7.2% (95%CI:6.72%-7.74%);the detection rate of 3D-OCT was significantly higher than routine examination of fundus for macular disease (χ2=79.05, P<0.01).Female, over 65 years old, surgical history of diseased eye, and high myopia were risk factors of cataract combined with macular disease, the relative risk was 1.705 (95%CI:1.091,2.664), 1.893 (95%CI:1.219,2.939), 6.593 (95%CI:2.027,21.447) and 95%CI:5.130 (2.841,9.263) respectively, the risk of cataract combined with macular disease showed an increasing trend with rising age.CONCLUSION:In preoperative examination of cataract patients, 3D-OCT has higher sensitivity in the detection of macular disease, especially for women, over 65 years old, high myopia and surgical history of diseased eye, 3D-OCT can be used as a routine preoperative examination.

OBJECTIVETo study the adaptive response mechanisms in high background radiation area (HBRA) among Yangjiang local people through gene and protein expression of receptor for advanced glycation end products (RAGE) and S100A6 in peripheral blood and sputum in inhabitants of HBRA.

METHODSA total of 53 male inhabitants were selected from HBRA in Yangjiang as the exposure group, while 53 male inhabitants were selected from Enping (control area, CA)as the control group. The content of RAGE and S100A6 gene and protein were detected by RT-PCR and Western blotting assay. Thermo luminescent dosemeter(TLD) assay was used to measure the outside dose and estimate the effective dose.

RESULTSThe effective dose in CA and HBRA was respectively 1.95 mSv and 6.24 mSv, which was 3 fold difference. Compared with CA, RAGE and S100A6 expression were significantly reduced in both gene and protein level in HBRA. The relative median mRNA expression of RAGE and S100A6 in peripheral blood were respectively 0.28, 1.06 and 0.16, 0.79 in CA and HBRA group, there was significance (with analysis Z values of -2.587 and -2.328 respectively, P < 0.05) with Wilcoxon rank test. For the protein of sputum, the relative median expression were respectively 2.98, 2.25 and 0.53, 0.47 with significant difference (with analysis Z values of -2.201 and -2.366 respectively, P < 0.05) by Wilcoxon rank test.

CONCLUSIONThe low expression of RAGE and S100A6 in HBRA group might be correlated with the adaptive response and the low mortality of cancer in HBRA.

Adaptation, Physiological ; radiation effects ; Background Radiation ; Cell Cycle Proteins ; metabolism ; China ; Humans ; Male ; Middle Aged ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; S100 Calcium Binding Protein A6 ; S100 Proteins ; metabolism ; Sickness Impact Profile

Objective: To evaluate the effects of dietary behaviors on the risk of hypertension, diabetes and cardiovascular diseases. Methods: A total of 12 208 subjects aged 18-60 years old were investigated by questionnaires to collect demographic data, dietary behaviors and lifestyle information, when they did health examination in a tertiary hospital in Beijing from 2014 to 2019. During the observation period of five year, the incidence of hypertension, diabetes and cardiovascular diseases were collected through health examination files every year. The multivariate logistic regression model was employed to analyze the associations of dietary behaviors with hypertension, diabetes and cardiovascular diseases. Results: The study included 6 218 ( 50.93% ) males and 5 990 ( 49.07% ) females. The cumulative incidence rates of hypertension, diabetes and cardiovascular diseases were 7.75%, 2.72% and 3.49%, respectively. The multivariate logistic regression analysis indicated that the high-sodium diet ( OR=1.422, 95%CI: 1.191-1.697 ) , eating fast ( OR=1.457, 95%CI: 1.102-1.974 ), eating more refined grain ( OR=1.251, 95%CI: 1.050-1.490 ) and drinking milk less than once a week ( OR=1.316, 95%CI: 1.022-1.697 ) were risk factors for hypertension. The high-sodium diet ( OR=1.344, 95%CI: 1.048-1.725 ), eating fast ( OR=1.733, 95%CI: 1.046-2.871 ), eating more meat ( OR=1.651,95%CI: 1.263-2.158 ) were risk factors for diabetes. High-sodium diet ( OR=1.501, 95%CI: 1.192-1.889 ) was risk factors for cardiovascular disease. Conclusion The diet with high sodium, more meat and refined grain as well as eating fast can increase the risk of hypertension, diabetes and cardiovascular diseases.

OBJECTIVE: To profile urinary metabolite variations from 1, 2-dimethylhydrazine (DMH)-induced precancerous colon rats, Jinfu Kang treated rats and healthy controls.

METHODWe used ethyl chloroformate derivatization and gas chromatography-mass spectrometry (GC-MS) based metabonomic method to analyze rat urines.

RESULTThe time-dependent variations of metabolite profile showed a progressive deviation of the metabolism in the model group from the initial pattern over time and a systemic recovery of the metabolism in the treatment group, which is consistent with the histological results. The in-depth analysis indicated that the disorder of tricarboxylic acid cycle (TCA), tryptophan metabolism, polyamine metabolism and gut flora structure were associated with DMH intervention.

CONCLUSIONMetabolic study revealed that Jinfu Kang can effectively reverse metabolic departures in DMH-induced precancerous colon rat, which is consistent with pathological results.

Animals ; Colonic Neoplasms ; chemically induced ; pathology ; Colonic Polyps ; chemically induced ; drug therapy ; urine ; Dimethylhydrazines ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Gas Chromatography-Mass Spectrometry ; Male ; Rats ; Rats, Wistar

OBJECTIVETo establish a chiro chromatography for studying the stereoselective metabolism of propranolol (PL) in S(9) incubates prepared from transgenic cell lines expressing human cytochrome P450.

METHODSThe concentration of each enantiomer in S(9) incubates was determined through precolumn derivatization with GITC, followed by RP-HPLC assay using S-(+)-propafenone as internal standard.

RESULTSBaseline separations among the diastereomers of S(-)-P, internal standard and R(+)-PL were achieved on Shimpack CLC C(18)ODS column, with UV detection and methanol:water:glacial acetic acid (67/33/0.05,v/v/v) as mobile phase. The assay was simple, accurate, precise and specific. The linear range was from 5 to 500 micromol/L for each enantiomer. The limit of quantitation (LOQ) for the method was 5 micromol/L for the S(-)-and R(+)-PL, respectively (n=5, RSD<10%). The analytical method afforded average recoveries of 98.7 and 98.1% for S(-)- and R(+)-PL, respectively. The reproducibility of the assay was good (RSD<10%). The time-dependent studies showed that PL had the stereoselectivity of S-(-)-isomer in metabolism via CYP2C18 and the stereoselectivity of R-(+)-isomer in metabolism via CYP2C9.

CONCLUSIONThe method allows to study of stereoselective metabolism of PL in vitro.

Chromatography, High Pressure Liquid ; Cytochrome P-450 Enzyme System ; genetics ; physiology ; Humans ; Propranolol ; analysis ; metabolism ; Reproducibility of Results ; Stereoisomerism ; Transgenes

OBJECTIVETo investigate the effect of Rongshi granule on osteopontin(OPN) expression.

METHODThe urlisthiasis rats were induced by ethylene glycol (EG) and ammonium chloride, the control group rats were non-treated, and the Rongshi granule groups (low-dose group, middle-dose group and high-dose group) were administered Rongshi granule in addition to EG and ammonium chloride in 21 days. Pooled 24 h urine samples from each group were collected weekly with the use of metabolic cages, the concentration of uric calcium and oxalic acid were respectively measured by EDTA and photoelectric colorimetric method. Eight animals from each group were killed at 0, 7, 14, and 21 days, kidneys were histologic examinaed and immunohistochemical staining.

RESULTThe expression of kidney osteopontin in model group was obviously higher than that of control group (P <0.01), and was up to the highest at 21 days with 1.4 times (0.281 3/0.201 8) of the control group. The expression of kidney osteopontin in all of the Rongshi granule groups were lower than those of model group (P < 0.05), with an obvious dose-dependent manner. The degree of the kidney calcium oxalate crystal of the rats in all the Rongshi granule groups was much lower than that of model group, and the uric calcium and oxalic acid were much lower than those of model group (P < 0.01).

CONCLUSIONThe Rongshi granule could inhibit the expression of osteopontin in rat urolithiasis model.

Ammonium Chloride ; Animals ; Calcium ; urine ; Calcium Oxalate ; metabolism ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Ethylene Glycol ; Female ; Kidney ; metabolism ; Kidney Calculi ; chemically induced ; metabolism ; Male ; Osteopontin ; metabolism ; Oxalic Acid ; urine ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley