The desired DNA product of KGPcd and KGP-hag was obtained from the total DNA of Porphyromonas gingivalis by PCR with two pairs of gene specific primers. The segment of KGPcd and KGP-hag (about 1.5kb and 1.6kb) was inserted into pGEM-T easy Vector. The double-stranded DNA of the postitive clone was analyzed by restriction endonuclease mapping and DNA sequenceing. The sequences of KGPcd and KGP-hag were consistent with those of the references appeared. The proteins of KGPcd and KGP-hag will be obtained for further study.

Objective To compare clinical features,pulmonary functions,chest imaging and prognosis between combined pulmonary fibrosis and emphysema syndrome (CPFE) and idiopathic pulmonary fibrosis(IPF) without emphysematous changes in elderly patients.Methods 88 elderly IPF patients in Beijing hospital from January 2000 to October 2012 were divided into CPFE (n=30) and IPF (n =58) groups according to the CT imaging.Clinical features,blood gas analysis,pulmonary function,chest CT and survival time were compared between the two groups.Results 30 CPFE patients with the mean age of(75.5 ±7.6) years and 58 IPF patients without emphysema with the mean age of(73.7±6.8) years were enrolled.The proportions of male patients,smoking history and mortality were higher in CPFE patients than in IPF group(86.7％ vs.63.8％,28 vs.36,76.7％ vs.43.1％,x2 =5.09,9.74,8.98,P＜0.05 or 0.01).CPFE patients had a higher force vital capacity(FVC) and total lung capacity(TLC) as compared with IPF group [(2.6±0.9) L vs.(2.1± 0.5) L,(5.4±1.9) L vs.(4.4±1.1) L,t=2.69,2.35,P＜0.01 or 0.05].There were no significant differences in forced expiratory volume in one second(FEV1) and the diffusion capacity for carbon monoxide(DLCO) between the two groups.The main type of emphysema by HRCT scan were centrilobular emphysema in CPFE patients.There were lower median survival time in CPFE group than in IPF group [(3.0±0.2) years vs.(4.0±1.0) years,x2=4.50,P＜0.05].Conclusions The majority of elderly CPFE patients are males and smokers.The lung volume is increased in elderly CPFE patients as compared with IPF patients.Centrilobular emphysema is the primary type in CPFE patients.The prognosis is worse in elderly CPFE patients than in IPF patients.

OBJECTIVE:To establish a method for simultaneous determination of 6 residual organic solvents in aprepitant raw material as methanol,ethanol,acetone,isopropyl alcohol,methyl tert-butyl ether and tetrahydrofuran.METHODS:Headspace capillary gas chromatography was adopted.The determination was performed on DB-624 capillary column using temperature programming.The temperature of injector port was 180 ℃,and flame ionization detector was used with temperature of 260 ℃.Nitrogen was used as carrier gas with flow rate 3.0 mL/min.The spilt ratio was 5 ∶ 1,and head-space injection volume was 1.0 mL.The head-space equilibrium temperature was set at 80 ℃,and equilibrium time was 40 min.RESULTS:The linear ranges of methanol,ethanol,acetone,isopropyl alcohol,methyl tert-butyl ether,tetrahydrofuran were 6.052-605.232 μ g/mL (r=0.999 9),9.987-998.718 μg/mL(r=0.999 9),9.998-999.768 μg/mL(r=0.999 8),9.986-998.634 μg/mL(r=0.999 9),9.991-999.090 μg/mL (r=0.999 7),1.461-146.133 μg/mL(r=0.999 5),respectively.The limits of quantitation were 1.782 1,2.079 0,0.749 8,1.777 8,0.223 1,0.607 0 μg/mL;the limits of detection were 0.594 0,0.693 0,0.249 9,0.592 6,0.074 4,0.202 3 μg/mL,respectively.RSD of precision test was lower than 2.0％.Only acetone and isopropyl alcohol were detected in stability test and reproducibility tests,RSD＜2.0％.Their recoveries were 99.34-100.75％ (RSD=0.52％,n=9),98.20％-100.24％ (RSD=0.69％,n=9),98.07％-100.07％ (RSD=0.84％,n=9),99.86％-101.32％ (RSD=0.58％,n=9),97.87％-104.02％ (RSD=2.13％,n=9),98.26 ％-100.58 ％ (RSD =0.75 ％,n =9),respectively.CONCLUSIONS:The established method is simple,accurate and reproducible,and can be used for simultaneous determination of 6 residual organic solvents in aprepitant raw material.

Objective: To develop ~(99)Tc~m labeled dopamine transporter(DAT) imaging agent ~(99)Tc~m-(2?-[N,N~(,)-bis(2-mercaptoethyl)ethylenediamino]methyl,3?-(4-chlorophenyl)tropane(TRODAT-1) for evaluating changes of DAT in patients with Parkinson disease(PD). Methods: The SD rats were divided into control group(n=5),PD models group(n=22)and generalized cerebral infarction models group(n=5).~() Unilateral smashing and injecting autothrombo into carotid artery of SD rats were used.~(99)Tc~m-TRODAT-1 distributing in normal rat striatum was observed.The uptakes in sound side and smashed side of PD rats striatum and in two sides of multiple infarction rats striatum were compared. Results:~(99)Tc~m-TRODAT-1 distribution in normal rats striatum exhibited a obvious uptake in striatum.And PD rats results exhibited that the uptake was less in normal striatum than in smashed striatum obviously.The result of multiple infarction rats is same as normals rats. Conclusion: ~(99)Tc~m-TRODAT-1 might betaked specificly.Its imaging can provide a beneficial evidence for PD disease early diagnose.

A study on ariboflavinosis was made among the students of a college in Chongqing from Jan. to Apr. 1980. The incidence of ariboflavinosis was 18.3%. There were typical oro-genital syndromes, e.g. cheilosis, angular stomatitis, glossitis and scrotal dermatitis etc. 84 students suffered fro-m 3 types of scrotal dermatitis, The incidences of squamo-erythematoid pa-puloid and eczematoid type were 7.26%, 19% and 8.3% respectively. A patient with scrotal dermatitis(eczematoid type) was accompanied with similar lesions on the skin of buttock, right chest and right upper arm and they subsided almost at the same time with scrotal lesion after riboflavin treatment.Ten subjects of scrotal dermatitis were selected for fungus examination and it gave negative findings both microscopically and culturally.Dietary survey was made in the student's dining hall and the nutrients were calculated. The amount of riboflavin averaged only 1.07 mg per capita per day, but the calculated value was probably higher than the actual intake due to improper cooking. The intakes of thiamine, niacin and ascorbic acid were adequate.The 4-hour urinary riboflavin excretions of 10 subjects following a 5mg oral load averaged 590?g.Dietary survey, biochemical test and clinical examination benefited the diagnosis of ariboflavinosis. The lack of the knowledge of nutrition and improper cooking of vegetables were perhaps the main causes of such deficiency.

The pancreatic enzyme-II type collagenase digestion method was adopted for primary culture of osteoblasts, inoculation and passage. They were identified by alkaline phosphatase dye-liquor. N-butanol extract fractions from different processed products of Cibotium barometz were prepared. The above osteoblasts were jointly cultured with protocatechuic acid, protocatechuic aldehyde, kojic acid and the mixed control liquid of the above three substances, and their proliferation was detected by CCK-8. Various n-butanol extract fractions from different processed products of C. barometz showed a significant proliferative effect on osteoblasts in the order of the wined > the heated > the salted > the sand-heated and wined system > the alcohol-processed > the steamed > the crude. The q test showed no significant difference among sand-heated, alcohol-processed and steamed C. barometz, no significant difference between heated and salted C. barometz. Various control substances also showed a certain proliferative effect on osteoblasts in the order of the mixed control > protocatechuic aldehyde > protocatechuic acid > kojic acid. The q test showed no significant difference between protocatechuic aldehyde and protocatechuic acid. All of n-butanol extract fractions from different processed products of C. barometz showed a significant effect on osteoblast proliferation, of which wined C. barometz showed the best effect. All of phenolic compounds such as protocatechuic aldehyde, protocatechuic acid and kojic acid showed a significant proliferative effect on osteoblasts.
Animals ; Cell Proliferation ; drug effects ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Osteoblasts ; cytology ; drug effects ; Pteridophyta ; chemistry ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of Smad7 on the expressions of collagen I and alpha-smooth muscle actin (alpha-SMA) in HSC-T6 cell line activated by transforming growth factor-beta1 (TGF-beta1).

METHODSHSC-T6 cells stably expressing M2-flag protein were selected after co-infection of the cells with pTRE-Smad7-M2-flag and pTet-on. The optimal dose of doxycycline for inducing Smad7 was determined, and the effects of Smad7 over-expression on the expressions of collagen I and alpha-SMA in the cells activated by TGF-beta1 and on Smad2/3 phosphorylation were evaluated using Western blotting.

RESULTSThe optimal dose of doxycycline for inducing Smad7 expression was 2 mg/L. Smad7 over-expression induced by doxycycline decreased the expressions of collagen I and alpha-SMA in HSC-T6 cells activated by TGF-beta1, and down-regulated the level of Smad2/3 phosphorylation.

CONCLUSIONSmad7 over-expression inhibits Smad2/3 phosphorylation, and decreases the expression of collagen I and alpha-SMA in HSC-T6 cells induced by TGF-beta1 to inhibit the progression of liver fibrosis.

Actins ; genetics ; metabolism ; Cells, Cultured ; Collagen Type I ; genetics ; metabolism ; Genetic Therapy ; Hepatocytes ; cytology ; metabolism ; Humans ; Liver Cirrhosis ; metabolism ; therapy ; Smad7 Protein ; pharmacology ; Transforming Growth Factor beta1 ; pharmacology

Objective To investigate the anti-atherosclerosis effect of apocynum venetum(AV)by observing the influence of AV extract on early inflammatory factor TNF-αexpression.Methods Human U937 monocytes were differentiated to macrophages by the phorbol myristate acetate(PMA)induction and acted with 100 mg/L ox-LDL to form the foam cells for establishing the early atherosclerosis model(ox-LDL group).The different concentrations(0.2,0.4,0.8 mg/L)of AV were added to co-culture for 48 h (AV1,AV2,AV3 groups).The expression level of TNF-αin the supernate was detected by ELISA and RT-PCR respectively.Re-sults Compared with control group,the expression level of TNF-αin the ox-LDL group was significantly increased,the expression level of TNF-αin various AV medication groups(AV1,AV2,AV3 groups)was significantly decreased compared with the ox-LDL group(P<0.05).The AV concentration increase was negatively correlated with the TNF-αexpression level(P<0.05).Conclusion TNF-αis an important inflammatory factor in early atherosclerosis,AV could play the anti-atherosclerosis role by inhibiting the inflammatory factors.

OBJECTIVETo observe the long-term apparent and structural change of the neo-urethra inner wall reconstructed with different kinds of tissue. To compare the difference between normal and reconstructed urethra for urethra reconstruction with more appropriate materials.

METHODSThe neo-urethra inner wall was observed during operation, through urography and urethroscope. The tissue section of neo-urethra inner wall was observed through light microscope and electron microscope.

RESULTSWith unchanged structure, the appearance of neo-urethra reconstructed with skin or mucosa could be close to normal urethra after a long time. Neo-urethra made of buccal mucosa possesses some of the most important basic structures for normal urethral microenvironment to form. The structure of neo-urethra made of bladder could be close to normal urethra after a long time.

CONCLUSIONSBuccal and bladder mucosa may be a better material than skin for urethra reconstruction.

Adolescent ; Adult ; Biocompatible Materials ; Child ; Female ; Graft Survival ; Humans ; Hypospadias ; surgery ; Male ; Mouth Mucosa ; transplantation ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Tissue Culture Techniques ; Tissue Engineering ; Tissue Transplantation ; methods ; Urethra ; surgery ; Young Adult

OBJECTIVEUsing polymerase chain reaction-reverse blot dot (PCR-RDB) technique to establish a new method for hepatitis C virus (HCV) genotyping and to study the distribution of HCV genotypes in Foshan area.

METHODSHCV primers and probes were designed in 5'-untranslated region (nt-1-nt-299) of HCV. HCV RNA in serum was isolated and purified, and its cDNA was obtained by reversed transcription. Nested PCR using biotin-labelled primers, was done. PCR products were hybridized with immobilized specific probes (genotype 1a to 3b) on Biodyne C membrane to genotype HCV by color development while adding POD and TMB. A certain judgment could be made according to the position of color reaction. The reliability of this new method was verified by sequencing. HCV RNA levels in serum were determined by real time fluorescent quantitative (FQ)-PCR. 60 FQ-PCR-positive HCV sera from Foshan area were genotyped using this assay.

RESULTSAll 60 sera could be successfully genotyped by PCR-RBD. 50 (83.3%) cases were found to be genotype 1b, 2 (3.3%) as genotype 1a and 2 (3.3%) as genotype 2a while 5 (8.0%) to be mixture of genotype 1a and 1b, and 1 (1.7%) to be mixture of genotypes 1b and 2a. No genotypes 2b, 3a and 3b were found. The results of PCR-RDB genotyping methods coincided with sequence analysis.

CONCLUSIONNewly established HCV genotyping system was proved to be sensitive, specific, precise and economic, thus suitable for clinical and epidemiologic studies. The results of HCV genotyping showed that genotype 1b was the predominant genotype in Foshan area.

Genotype ; Hepacivirus ; classification ; genetics ; Hepatitis C ; virology ; Humans ; Immunoblotting ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity