Adult ; Female ; Gastritis ; therapy ; Humans ; Male ; Massage ; Middle Aged

AIM: To investigate the effect of compound rhubarb preparation (Kintop) in preventing obesity in rats and its probable mechanism involved. METHODS: Twenty-six newborn SD rats were randomly grouped as rhubarb preparation plus high-energy forage group( n= 8), high-energy forage control group( n= 8) and ordinary forage control group( n= 10). The rats in rhubarb preparation plus high-energy forage group and high-energy forage control group were fed with high-energy forage and those in ordinary forage control group were fed with ordinary forage. The rats in rhubarb preparation plus high-energy forage group were administered by compound rhubarb preparation (40 mg?100 g -1 body weight?d -1 ) from 9th to 17th week. The dynamic changes in body weight, celiac fat weight and adipocytes size were measured. Immunohistochemical analysis of leptin in celiac adipocytes (ABC method) and measurement of serum leptin level (RID method) were performed. RESULTS: The body weight and the wet weights of celiac fat were lower, their adipocytes were smaller and immunohistochemical stainings of leptin were weaker in rhubarb preparation plus high-energy forage group than those in high-energy forage control group. There was an obvious positive correlation between the expression of leptin and celiac fat tissue weight( r= 0.8663, P

The copy numbers of exogenous gene in transgenic animals is always regarded as an important information of transgenic animals.Thus,simple and sensitive methods are required for the detection of the copy numbers of exogenous gene.Three kinds of transgenic Shanbei white cashmere goats,containing Tβ4-GFP,FGF5s-GFP and VEGF164-GFP,has been obtained by using PiggyBac(PB) transposon system.Fluorescence quantitative PCR was carried out to detect the copy numbers of copGFP.Using Gluc as reference gene,the double standard curves of exogenous gene and reference gene were mapped and the genomic DNA of transgenic goats were analysized by real-time fluorescence quantitative PCR.Moreover,the copGFP/Gluc ratio in the samples was calculated as the copy numbers of copGFP.In addition,Tβ4-GFP transgenic cashmere goats were selected to detect the integration sites by using the genomic walking kit.The results showed that the standard curve equation of copGFP was y=-3.230 6x+39.216 (R2 =0.998 8) and the standard curve equation of Gluc was y=-3.564 8x+38.440 (R2 =0.996 0).The copy numbers of exogenous gene in the transgenic cashmere goats were obtained and the numbers of integration sites in the selected Tβ4-GFP transgenic goats were consistent with the copy numbers of copGFP.As a conclusion,the high throughput,fast and sensitive real-time fluorescence quantitative PCR is an efficient and convenient method for the copy number of exogenous gene in transgenic cashmere goats.

OBJECTIVETo investigate the distribution character of the mutations of 6-pyruvoyl tetrahydropterin synthase (PTPS) gene and to provide effective basis for gene diagnosis of tetrahydrobiopterin deficiency (BH4D) in children with hyperphenylalaninemia.

METHODSDirect sequencing was performed for screening the PTPS gene mutations in 5 patients with clinically suspected BH4D and their parents. The nature of the novel mutations were deduced by the sequences alignment and the structural analysis of mutant protein. Artificial construct restriction site was used to detect the novel mutation in the control samples. The dates of urinary pterin analysis and BH4 loading test were retrospectively analyzed after gene analysis.

RESULTSFour PTPS gene mutations (N52S, P87S, D96N, and L127F) were detected in our study. The genotypes of four PTPS deficiency patients were identified as N52S/L127F, P87S/D96N, N52S/D96N, and D96N/ -. As a novel mutation that has not been reported previously, the mutation L127F was not detected in 50 normal controls. This novel mutation L127F was inherited from the patient's mother, and this mutant site was highly conserved by sequences alignment and the protein structural analysis. Four of the five cases with hyperphenylalaninemia and suspicious BH4D, whose urinary biopterin percentage was lower than 2% , were diagnosed as PTPS deficiency during 5-20 months old. The remaining one case was excluded from BH4D.

CONCLUSIONSThe mutant characterization of PTPS gene was coincident with other early studies in Chinese. The novel mutation L127F was considered as a pathogenetic mutation and associated with severe clinical phenotype.

Asian Continental Ancestry Group ; genetics ; Biopterin ; analogs & derivatives ; deficiency ; genetics ; Child, Preschool ; DNA Mutational Analysis ; methods ; Humans ; Infant ; Infant, Newborn ; Mutation ; genetics ; Phenylketonurias ; genetics ; Phosphorus-Oxygen Lyases ; genetics ; Polymerase Chain Reaction

A HPLC-MS/MS multiple-reaction monitoring (MRM) quantitative analysis was made to establish a determination method for drug concentrations of costunolide (Co) and dehydrocostuslactone (De) in blood samples in the positive ion mode, with diazepam as the internal standard substance, in order to study the pharmacokinetic process of sesquiterpene lactones costunolide and dehydrocostuslactone after the oral administration of Weichang'an pills, and provide an theoretical basis for further studies on the substance basis for the anti-diarrhea effect of Weichang'an pills. In the blood samples, Co and De showed a good linearity within concentration ranges 0.700 0-769.7, 2.510-956.0 μg x L(-1), respectively. The results of precision, stability and recovery experiences proved the stability and reliability of the plasma concentration determination method. After the oral administration, the concentrations of Co and De in plasma increased with the increase in dose, with T(max) between 10.65-12.98 h, indicating a long time to reach peak plasma concentrations; C(max) of costunolide and dehydrocostuslactone ranged between 3.750-5.450,15.34-44.52 μg x L(-1), respectively. The in vivo adsorption of Co and De conformed to the one-compartment model, with a longer time to attain the peak plasma concentrations. These results provided an experimental basis for revealing the active substance basis and clinical medication of Weichang'an pills.
Administration, Oral ; Animals ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; pharmacokinetics ; Lactones ; administration & dosage ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Wistar ; Sesquiterpenes ; administration & dosage ; blood ; pharmacokinetics ; Tablets ; administration & dosage ; chemistry ; pharmacokinetics

Objective To examine the induction effects of bone marrow mesenchymal stem cells(BMSCs) transfected with bone morphogenetic protein 7 (BMP7) gene differentiating into chondrocytes. Methods We observed the phenotype of cells which were stained with alcian blue and HE climbing to the six pore plate with invert microscope. The glycosaminoglycan (GAG) value in culture medium was detected in control group,BMP7 transfect and culture medium induced groups after 7,14 and 21 days using standard curve method. Standard curve was described using galacturonic-acid as reference substance. The content of collagen Ⅱ was detected by ELISA method. Results HE and Alcian blue staining showed that BMP7 gene transfection group and the group induced by fluid possess the characteristics of chondrocyte. BMP7 induced BMSCs differentiation to chondrocyte which secrete specific protein called collagen Ⅱ and GAG. Content of GAG were (17.1±3.4),(39.5±5.4),(40.8±6.1)mg/L in control group,BMP7 gene transfected group and induced group,collagen Ⅱ were (89.7±14.3),(152.8±14.5),(155.5± 19.3)μg/L in these three groups separately. Comparing with control group,GAG and collagen Ⅱ of BMP7 gene transfected group and culture medium induced group increased obviously(all P < 0.05),but there was no significant difference between BMP7 gene transfeeted group and culture medium induced group (P > 0.05). Conclusion This active protein induces BMSCs differentiating into chondrocyte,in a level similar to that of inducing medium.

Objective To identify the effect of both high glucose and high insulin on miR-145 level.Methods The human vascular smooth muscle cells ( VSMCs) were cultured and the proliferation of VSMCs was induced by high glucose and high insulin medium.The samples were divided into 4 groups: normal group, high glu-cose group (25 mmol/L), high insulin group (300 mU/L), high glucose and high insulin group.The ex-pression of miR-145 in VSMCs was assayed by real-time PCR.Proliferation of VSMCs was determined by MTT method After 72 h cultivation.The migration of VSMCs was analyzed by cell scratch test .VSMCs in each group was transfected by miR-145 virus ( lentiviral vector ) .Proliferation and migration were assayed after 48 h transfection.Results The expression of miR-145 in VSMCs of other three groups was decreased ( P<0.05 ) , especially the expression in the high glucose and insulin group was the lowest ( P<0.01 ) .Prolifera-tion and migration of VSMCs was promoted by high glucose and/or high insulin medium.Under fluorescent, transfection rate of VSMCs was about 80%after 48 h transfection.Proliferation and migration of VSMCs in each group after transfection were significantly lower than before ( P<0.05 ) .Conclusions High glucose and high in-sulin could decrease the expression of miR-145 in VSMCs, The overexpression of miR-145 may inhibit the prolif-eration and migration of VSMCs .

Objective To evaluate the triage value of high-risk human papillomavirus (HR-HPV)detection in women with atypical squamous cells of undetermined significance (ASCUS).Methods Two huandred women with atypical squamous cells of undetermined significance (ASCUS) were selected by thinprep cell test(TCT) simultaneously using HPV DNA testing by hybrid capture Ⅱ (HC) and biopsy under the colposcopy,compared the HR-HPV DNA testing with final pathological diagnosis.CIN (cervical intraepithelial neoplasia) means cervical biopsy positive.Results The positive rate of cervical biopsy in HPV positive patients was 60.38％(96/159),and in HPV negative patients was 21.95％(9/41),the difference was obviously (P ＜ 0.05).Conclusion HR-HPV detection is a good triage approach for the patients with ASCUS.

Objective To evaluate the clinical value of fiberoptic ductoscopy in diagnosis and treatment of patients with galactophoritis or mammary duct ectasia. Methods From November 2005 to March 2008, fiberoptic ductoscopy were performed in 120 women with nipple discharge. The duct of 95 cases as non-malignant lesion were insufflated and perfusioned with entamycin and dexamethasone. Results Ninty-five of 120 cases were non-malignant disease,which contained one side 81 and two sides 14; the discharge was bloody,ivory, stramineous in 21, 17, 57 patinents, respectively; and the dignosis were 17 mammary duct ectasia, 53 galactophoritis, and 25 mammary duct ectasia with galactophoritis. Of the 95 cases, hich were intradutal insufflated and perfusioned with gentamycin and dexamethasone, the nipple discharge were decreased or disappeared in 81 cases, the effective rate was 85.3%. Conclusion Fiberoptic ductoscopy is a convenient,safe, accurate method in diagnosis and treatment of patients with galactophoritis or mammary duct ectasia.