Objective To establish a method for the determination of Baicalin in Fuke Yangkun pills.Method High-performance liquid chromatography method was used.The chromatographic conditions were as follows:C18 column,mobile phase of methanol ∶0.4 %phosphoric acid(45 ∶55),detection wavelength being 280 nm,a flow rate of 1.20 mL?min-1 and the column temperature being 25 ℃.Results There existed a good linearity when the injection quantity is in the range of 0.0541 ?g to 0.5408 ?g.Average recovery is 99.98 %,and RSD is 1.32 %.Conclusion HPLC method is simple and effective and can be applied for the detemination of Baicalin in Fuke Yangkun pills.

Adolescent ; Female ; Humans ; Jaw ; pathology ; Parathyroid Neoplasms ; complications ; pathology

This study was aimed to explore five Chinese medicine components or ingredients from Qu-Shi Hua-Y u Decoction (QSHYD) with uniform design method and screen ingredients assembling of Chinese medicine BZL pre-scription in order to verify its therapeutic effect on experimental fatty liver rats. High-fat diet was used in the estab-lishment of fatty liver rat models. Five effective ingredients (i.e., chlorogenic acid, geniposide, curcumin, polydatin and polysaccharide of Atractylodes macrocephala Koidz) of QSHYD were used as study subjects. Uniform design was applied in the grouping design. The intragastric administration was given for four weeks. The screening index was the content of liver triglyceride (TG). The ingredients assembling of Chinese medicine BZL prescription was screened through uniform design and regression analysis. The same fatty liver animal model was used in the comparison be-tween QSHYD and rosiglitazone. The TG content in liver tissues of rats and serum ALT activity were detected. The pathological changes of liver tissues were observed (HE stain and oil red O stain) with Ridit analysis to verify its therapeutic effect. The results showed that through screening, the regression equation was Y = 15.083X1 + 5.321X2- 5.186X3 - 16.157X4 + 9.35X5 + 17.667X3X4 - 8.422X1X2 - 6.617X3X5 + 16.571 (X1: chlorogenic acid, X2:polydatin, X3: polysaccharide of Atractylodes macrocephala Koidz, X4: geniposide, X5: curcumin). According to this equation, the best TG reducing effect occurred when the chlorogenic acid (X1), polysaccharide of Atractylodes macrocephala Koidz (X3), and geniposide (X4) were the maximum dosage. It showed that the best combination in inhibiting TG content in liver tissues was X3, X4 and X1 (i.e. BLZ prescription). Among fatty liver rat mod-els induced with same high-fat diet, the screened Chinese medicine ingredient BZL prescription can obviously reduce its TG content in liver tissues and serum ALT activity ( P < 0 . 01 ) . It obviously improved the fatty degen-eration of rat's liver. It was concluded that ingredients assembling of Chinese medicine BZL prescription screened through uniform design can obviously prevent and treat fat deposition and injury of the liver among rat models induced by high-fat diet .

The optimization on liquid fermentation and purification of the fibrinolytic enzyme from Stenotrophomonas maltophilia(DR-929) was investigated.The results showed the best fermentation are amidulin 2.0%,soya flour 1.0%,yeast extract 0.5%,NaCl 1.0%,CaCl2 0.02%,MgSO4 0.05%,inoculum of 36 hours,fermental time 4d,initial pH 8.0 or 9.0,temperature 25℃,volume of media 30ml,volume of inoculum 5% or 6%.The purification process includes the following steps: removing cells by the centrifugation,25%~70% saturation ammonium sulfate precipitation,HIC with Phenyl FF(high sub),IEC with Q-Sepharose FF,gel filtration chromatography with Superdex 75.SDS-PAGE electrophoresis was used to examine the purification effect,and the results indicated that homogeneous strap in SDS-PAGE and has a molecular weight around 28.3kDa.The purification factor and activity recovery of the fibrinolytic enzyme are 271.5 and 24.5%,respectively.

The specific fragment of Pneumococcal surface protein A(PspA)and Pneumococcal Surface Adhesin A(PsaA)gene was amplified by PCR from Streptococcus pneumonia 5 and Streptococcus pneumonia 19.The amplified fragnent of PspA and PsaA gene was ligated into pET-27b(+)vector and transformed into BL 21 E.coli for expression and obtain the expressive production of PspA and PsaA.Induced by IPTG,the expression level was as high as 75 % of the total disolube protein.The result showed that the recombinant plasmid could express a specific 75 kDa and 37 kDa fusion protein in E.coli BL 21,which showed the good immunogenicity and a broadly cross reactivity with the other serotypes.

Objective To observe the effect of S-adenosyl-L-methionine(SAMe) in treatment of jaundice in newborns and its mechanism.Methods Two hundred and two newborn infants with jaundice were treated with SAMe,76 cases in control group treated with phototherapy and liver enzyme induction elixir;SAMe 30-60 mg/(kg?d) were added to 202 cases intravenously in treatment group.The total biliorubin(T-BILI),direct bilinrubin(D-BILI) and indirect bilinrubin(I-BILI) were dynamically detected.Results Six days after treatment,the skin jaundice index in treatment group decreased remarkably.T-BILI,D-BILI and I-BILI decreased significantly.The curing effectiveness was higher in treatment group than that in control group.The number of applicating blood products and albumin,and blood produets/albumin were decreased in treatment group than those in control group.In those who used glucose to dissolve the SAMe 2.68% had blood-vessel phlebitis.Conclusions SAMe can efficiently quicken the retrogression of jaundice in newborns.It can reduce the use of blood products.It is a reliable and safe drug to treat jaundice in newborns.

OBJECTIVETo observe the effect of CKJ Recipe (consisting of Cordyceps sinensis polysaccharide, amygdaloside, and gypenosides) containing serum on the activation of rat primary hepatic stellate cells (rHSCs) and to explore its pharmacological mechanism.

METHODSrHSCs were isolated form liver and cultured for four days. Then they were divided into the normal control group, the model group, and the CKJ group. rHSCs in the model group and the CKJ group were treated with 2.5 ng/mL transforming growth factor beta1 (TGF-beta1) in serum-free DMEM for 24 h. Serum free DMEM (containing no TGF-beta1) was taken as the control for the normal control group. rHSCs in the CKJ group were treated with 5% CKJ-containing serum for 24 h. rHSCs in the other two groups were treated with 5% blank serum for 24 h.The protein expression level of a smooth muscle actin (alpha-SMA) was determined using high throughput screening (HCS) and Western blot. mRNA expression levels of alpha-SMA, collagen I (Col-I), platelet-derived growth factor receptor beta (PDGF-betaR), TGF-beta1, transforming growth factor beta receptor 1 (TGF-betaR1), and transforming growth factor beta receptor 2 (TGF-beta R2) were detected using quantitative RT-PCR.

RESULTSCompared with the normal control group, the protein expression level of alpha-SMA, mRNA expression levels of alpha-SMA, Col-I, PDGF-betaR, TGF-beta1, TGF-betaR1, and TGF-betaR2 significantly increased in the model group (P<0.05, P<0.01). Compared with the model group, the protein expression level of alpha-SMA, mRNA expression levels of alpha-SMA, Col-I, PDGF-betaR, TGF-beta1, TGF-beta1, and TGF-beta R2 significantly decreased in the CKJ group (P<0.05, P<0.01).

CONCLUSIONCKJ containing serum could inhibit the protein expression level of o-SMA, which was probably related with inhibiting TGF-beta1 and its related receptors.

Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hepatic Stellate Cells ; metabolism ; Rats ; Transforming Growth Factor beta ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVEExplore the genotype of the epidemic Japanese encephalitis virus (JEV) in Yunnan Province from molecular level and to understand the molecular differences of the virus isolated from Yunnan at different time, locality and host.

METHODSThree-day suckling mice were inoculated with viruses continuously and when the disease developed and the mice were dying, the brain was taken and DNA was extracted from the supernatants of the brain after grinding; then the gene fragments of Prm-C region were amplified by RT-PCR. The viral gene sequences were compared with those of other 72 strains of JEV originated from both China and abroad at different times. Finally the genotypes were analyzed with the method which was established by Woan-Ru Chen.

RESULTSAll the 3-day suckling mice which were inoculated with the virus died within 78 h. The results of the nucleic acid-sequence analysis showed that 17 strains of the experimental virus belonged to genotype 1 and 2 strains belonged to genotype 3. The difference between genotype 1 and type 3 were more than 15%. While the difference between 17 strains of genotype 1 which were separated at different time, location and hosts were only 3.8%-5.2%.

CONCLUSIONThe above results suggest that the genotype of the epidemic JEV in Yunnan Province are type 1 and 3 and the latter is the main type.

Animals ; Animals, Newborn ; Brain ; pathology ; virology ; China ; Encephalitis Virus, Japanese ; classification ; genetics ; isolation & purification ; Encephalitis, Japanese ; virology ; Genotype ; Mice ; Phylogeny ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Viral Envelope Proteins ; genetics

ObjectiveTo explore the prevalence and spectrum of β-thalassemia mutations in Fujian province,and to provide a reference for prenatal diagnosis and genetic counseling in this population.Methods Two thousand three hundred and one blood samples were randomly selected from 9 different areas of Fujian province from May 2008 to December 2010.PCR and reverse dot blot hybridization (RDB) were adopted for detection of the 17 common types of mutation,and the frequency of each genotype of β-thalassemia mutations was calculated.The β-globin gene of unknown positive samples were analyzed directly with DNA sequencing.Results Three hundred and fifty-nine cases were detected with β-thalassemia mutations of the 2301 copy blood samples submitted,and the detection rate was 15.60％ (359/2301).Of the mutated genes,12 different mutations were identified,namely IVS-2-654(C→T),CD41-42(-TCTT),CD17(A→T),-28(A→G),CD27-28(+C),CD26(G→A),CD71-72(+A),IVS-1-1(G→T),CD43(G→T),-29(A→G),initiation codon ATG→AGG and CD36(-C).Mutation frequencies were 46.54％ (175/376),33.24％ (125/376),9.31％ (35/376),5.05％ (19/376),2.13％(8/376),1.33％(5/376),0.80％(3/376),0.27％(1/376),0.27％(1/376),0.27％(1/376),0.53％(2/376),and 0.27％(1/376),respectively.The most common mutations were IVS-2-654 (C→T) and CD41-42 (-TCTT),which accounted for 79.78％(300/376) of total genetic mutations.In addition,a novel β-globin gene mutation CD36 (-C) allele was detected for the first time,the deletion of a nucleotide C at code 36 within exon 2 lead to a frameshift mutation that could result in a premature termination at code 60.Conclusions β-thalassemia mutations in Fujian province are complex with significant genetic heterogeneity.We present for the first time the detection of a new β-thalassemia mutation in the population:CD36(-C),which provides valuable information for genetic counseling and prenatal diagnosis in Fujian province.

Adolescent ; Adult ; Child ; Female ; Fracture Fixation, Intramedullary ; methods ; Humans ; Male ; Radius Fractures ; surgery