Objective:To optimize the extraction process of Hegan Lidan granules. Methods:In order to choose the optimal tech-nological parameters, the content of baicalin was determined by HPLC. An orthogonal method was utilized with solvent volume, extrac-tion time and extraction times as the impacting factors and the content of baicalin and extraction rate as the indices. Results:The opti-mal parameters were as follows:using 8-fold water as the solvent, the raw material was extracted three times with 2 h for each. Con-clusion:The process is steady and feasible, and can be used in the extraction of Hegan Lidan granules.

Objective To understand acute rejection differences between untreated recipients and rapmycin-treated recipients in a rat free flap allotransplantation model. Methods Brown groin free flaps were transplanted to Lewis recipients. In the treated group, recipients were treated with rapamycin at the dose of 4 mg/kg every day from day 0 to day 14 after transplantation. In the untreated group, recipients didn't receive any treatment. Allografts were evaluated clinically and histologically. Results Allografts in the treated group showed epidermolysis as sign of rejection.Rejection sign of untreated grafts was ischemic necrosis of whole skin. In histological evaluation, the treated grafts showed "band-like" lymphocytes infiltration in the upper dermis when rejection occurred, while the untreated grafts showed thrombosis in the subdermal vessels. Conclusion The differences between the two groups implied that embolization may be responsible for the rejection of free flap allotransplantation in rat model.

Objective: To establish the quality standard for compound Heishen pills. Methods: Scrophulariae Radix, Radix et Rhizoma and Belamcancae Rhizoma were identified by TLC. HPLC was used to determine the content of harpagoside and cinnamic acid in Scrophulariae Radix on a Welchrom-C18 column using methanol-acetonitrile-1% ethylic acid (8∶21∶71) as the mobile phase. The flow rate was 1. 0 ml·min-1 . The column temperature was at 30℃ and the detection wavelength was set at 278 nm. Results:The TLC method had good specificity without interference from negative control. The linear range of harpagoside was 1. 32-65. 80μg·ml-1 with the average recovery of 98. 06%(RSD=2. 16%),and that of cinnamic acid was 0. 38-19. 20 μg·ml-1 with the average recovery of 98. 78%(RSD=1. 34%). RSDs of precision, stability and reproductibility tests were all below 2%. Conclusion: The established method is accurate, feasible and reproducible. It can be used in the quality control of compound Heishen pills.

Objective To find out prevelence of thyroid goiter on school-aged children in excessive iodine region of Shandong Province.Methods By employing B-ultrasonography and palpation method,the thyroid volume were investigated and the thyroid goiter rate were calculated among 6415 children aged 8～10 years old in execssire iodine region.Results By employing B-ultrasonography method.The volumes of thyroid in goiter children aged 8,9,10 years were(5.67±1.49),(6.07±1.24),(7.30±2.01)ml,respectively.The goiter rates were 28.84%(683/2368),20.89%(448/2144),11.82%(225/1903).The volumes of thyroid in non-goiter children aged 8,9,10 years were(3.36±0.67),(3.64±0.77),(4.02±0.94)ml.When the normal value of thyroid volume in 8 and 9 year-groups lowered 0.5 ml,the goiter rate would be changed to 18.20%(430/2368)and 12.92%(277/2144) respectively.The thyroid volumes of boys and girls were(4.20±1.38),(4.18±1.73)ml,respectively.The goiter rates of boys and girls were 21.92%(732/3340)and 20.29%(624/3075)respectively.The rates of goiter detected by palpation in 8,9 and 10 year-groups were 10.05%(238/2368),10.31%(221/2144)and 14.45%(275/1903) respectively.Conclusions Along with the increasing age,the volume of thyroid in children aged 8～10 is increasing in excessive iodine region of Shandong Province.But the goiter rate is decreasing ahmg the increasing age.This phenomena may be caused by improper normal value of thyroid volume.Therefore,the normal value of thyroid volume should be revised urgently.

Aim To investigate the effect of Jinlida on cholesterol-related genes in skeletal muscle in fat-in-duced insulin resistance ApoE-/ - mice. Methods Ten male C57 BL/6 J mice were selected as normal group ( NF );50 male ApoE-/ - mice with a high-fat feeding after 16 weeks ( HF) were divided into model group, rosiglitazone ( LGLT ) , Jinlida low dose group ( JLDL, 0. 95 g · kg-1 · d-1 ) , Jinlida medium dose group ( JLDM, 1. 9 g·kg-1 ·d-1 ) , Jinlida high dose group (JLDH, 3. 8 g·kg-1·d-1), which were per-formed intragastric administration for 8 weeks. Oil red O staining of mouse skeletal muscle was used for fat ac-cumulation. Insulin receptor ( INSR) , insulin receptor body substrate-1 ( IRS-1 ) , low-density lipoprotein re-ceptor ( LDLR ) , cholesterol sensor ( SCAP ) mRNA and protein expression in mouse skeletal muscle were measured by quantitative reverse transcription PCR ( RT-PCR ) and Western blot. Results Compared with NF group, fasting blood glucose ( FBG) , choles-terol ( TC ) , triglyceride ( TG ) and low density lipo-protein cholesterol ( LDL-C ) of HF mice were signifi-cantly elevated, while high-density lipoprotein ( HDL-C ) significantly decreased ( P < 0. 05 ) . Compared with HF group, Jinlida group could reduce to varying degrees FBG, TC, TG and LDL-C in mice, and in-crease HDL-C ( P <0. 05 ) . Jinlida could downgrade fasting serum insulin ( FINS ) level, and improve the insulin sensitive index ( ISI ) ( P < 0. 05 ) . Jinlida could obviously improve skeletal muscle fat accumula-tion of mice. Compared with NF group, skeletal mus-cle INSR, IRS-1, LDLR mRNA and protein levels of HF group were significantly decreased ( P <0. 05 ) , while SCAP mRNA and protein level increased signifi-cantly (P<0. 05). Compared with HF group, Jinlida could increase to varying degrees INSR, IRS-1, LDLR mRNA and protein levels ( P < 0. 05 ) , and lower SCAP mRNA and protein levels ( P<0. 05 ) . Conclu-sion Jinlida can alleviate fat-induced insulin resist-ance in ApoE-/ - mice through regulation of cholester-ol-related gene expression.

Objective:To obtain oligonucleotide aptamers which can specifically bind to Bacillus anthracis spores by in vitro selection protocol-SELEX (system evolution of ligands by exponential enrichment).Methods:An in vitro synthesized 78 mer random DNA library (≤1014-15types of different DNAs ) was subjected to 15 rounds of selection using SELEX method against spores of B.anthracis vaccine strain A.16R. Binding of the aptamers to spores was visualized by biotin-streptavidin-horseradish peroxidase system.Results:PCR amplification band pattern of the first round selection was different from that of the ninth round. The binding assay demonstrated that D absorbance at 450 nm of the fifteenth round pool increased 9 times as compared with that of the first round , and the D absorbance increased with the increment of aptamers′ quantity binding to spores. Conclusions: A set of aptamers with considerable binding affinity to B.anthracis spores were successfully selected from the initial random DNA pool.

Objective To detect the immunogenicity of VirB9,a protein of type Ⅳ secretion system of Brucella.Methods Full length VirB9 gene was cloned into plasmid pET32a and expressed in Escherichia (E.) coli BL21 (DE3).Expression of recombinant protein was induced by isopropyl beta-D-thiogalactopyranoside (IPTG) and the recombinant fusion protein was purified by affinity chromatography on Ni2+-conjugated chelateing sepharose.The purity of the purified protein was ascertained by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDSPAGE) and the concentration was measured by bicinchoninic acid (BCA) protein assay kit.Animal model was established by immunizing BAL B/c mice with live vaccine strain S19 of Brucella and the mice immunized with phosphate buffered saline (PBS) as control.The blood of immunized mice was acquired after 4 weeks.Antibody against VirB9 in S19 immunized mice was detected by Rose Bengal plate agglutination test and serum tube agglutination test; IgG antibody titers against VirB9 in immunized mice were determined by enzyme linked immunosorbent assay(ELISA).At the 35th day,the immunized mice and control mice were killed and spleens were collected.The splenocytes were harvested and stimulated with each of VirB9,concanvalin A(ConA) or medium in triplicate.Production of gamma interferon (IFN-γ) was determined by enzyme-linked immunospot assay (Elispot).Results The full length of VirB9 gene was cloned into pET32a.The recombinant VirB9 protein was expressed at 43 × 103 in relative molecular mass and the purity of the purified recombinant VirB9 protein was above 97％ in SDS-PAGE and the concentration was 1.6 g/L in BCA protein assay.The antibody of VirB9 was detected in all S19 immunized mice but not PBS immunized mice by Rose Bengal plate agglutination test.The antibody titer in all S19 immunized mice was ＞ 1 ∶ 800 or ＞ 1 ∶ 3200 by tube agglutination test and ELISA,respectively.Meanwhile,the protein stimulated stronger IFN-γresponse in immunized mice than that in the control mice(147 cells Vs 38 cells).Conclusion VirB9 can stimulate humoral and cellular immunity and it might be an appropriate target for developing subunit vaccine against Brucella.

AIM: Cystostomy is the traditionary method for detecting urodynamic indexes in mice, which de-stroys the continuity of the bladder, and there are significant differences between this method and the clinically used trans-urethral method.This study aims to develop an appropriate urethral catheter to investigate the advantages and application val-ue of transurethral method for urodynamic test.METHODS:A pediatric intravenous catheter was used for urethral catheter-ization on 8 female mice, and linked to connect the catheter to baroreceptor and micropump.The epidural catheter was also used as manometry tube.RESULTS:Using this method, the following urodynamic indicators has been successfully cap-tured:basal bladder pressure (BBP), bladder leak point pressure (BLPP), maximum voiding pressure (MVP), maxi-mum bladder capacity ( MBC ) , post-void residual urine volume ( PVR ) , voiding volume ( VV ) , efficiency of voiding ( EV) and bladder compliance ( BC) .CONCLUSION:This is the first successful simulation used in human body to a-chieve mouse urodynamic testing through the urethra catheter, which avoids the impact of cystostomy on urodynamics in mice, and the mice are able to keep long-term survival after tests for the follow-up molecular and genetic experiments.

Objective To quantify the expressions of collagen metabolic markers carboxy terminal propeptide of type I procollagen(PICP),nitrogen terminal propeptide of type I procollagen (PINP),nitrogen terminal propeptide of typeⅢprocollagen(PⅢNP),type I collagen carboxy terminal telopeptide (ICTP), matrix metalloproteinases(MMPs)and the tissue inhibitor of metalloproteinases(TIMPs)in the serum of atrial fibrillation patients by enzyme linked immunosorbent assay(ELISA),and to discuss the atrial structural remodeling during atrial fibrillation(AF).Methods 71 elderly patients were enrolled,24 patients had permanent AF,24 patients had paroxysmal AF,and 23 patients were in sinus rhythm.The serum levels of all markers were measured by ELISA. Results PICP was increased in permanent AF group versus the paroxysmal AF group and sinus rhythm group by 25.4%and 42.8%(all P<0.05),respectively.PⅢNP was increased in permanent AF group versus the paroxysmal AF group and sinus rhythm group by 17.9a% and 35.6%(all P<0.05),respectively,and was increased in the paroxysmal AF group versus the sinus rhythm group by 15.0%(P<0.05).PINP and ICTP did not differ significantly between the 3 groups(all P >0.05).MMP-1 was significantly increased by 25.6%(P<0.05)in the paroxysmal AF group versus the sinus rhythm group.MMP-2 was also significantly increased in permanent AF group versus the paroxysmal AF group and sinus rhythm group by 54.9%and 37.9%(all P<0.05),respectively.MMP-7,MMp-9 and TIMP-1 did not differ significantly between the 3 groups(P>0.05).TIMP-2was significantly decreased in the permanent AF group and paroxysmal AF group versus the sinus rhythm group by 21.8%and 11.8%(P<0.05),respectively. Conclusions Disturbance in the balance of MMP/TIMP system may perturb the balance of collagen synthesis and degradation during atrial fibrillation.This may be a contributing mechanism to atrial structural remodeling in atrial fibrillation,and may correlate with the initiation and maintenance of AF.

Objective To compare the differences of lung pathological changes of acute lung injury in mice in-duced by lipopolysaccharide ( LPS) and graphite particles, and to explore the possible mechanisms of acute lung injury in-duced by fine particles of different origins.Methods 140 male specific-pathogen-free Kunming mice weighing 18-20 g were randomly divided into 7 experimental groups, in addition to the normal control group.The experimental groups were treated by intratracheal instillation of LPS solution or graphite powder suspension in different doses, respectively, to induce acute lung injury in the mice.The mortality of the mice was observed, and pathological changes of the lung tissues were ex-amined by light and transmission electron microscopy.Western blot was used to detect the protein expression of neutrophil elastase ( NE) in lung tissues , and real-time quantitative PCR was used to detect mRNA expression of monocyte chemotac-tic protein-1 ( MCP-1) in the lung tissue .Results Compared with the normal control group, some pathological changes were observed in the lung tissues of the groups L ( LPS) and G ( graphite) .There were numerous macrophages in the lung tissues in the group G mice, and exudate, mainly neutrophils, in the lung tissues of the group L.The NE protein expres-sion in the lung tissue was significantly higher than that of the normal control group ( P<0.05) , and there was also a sig-nificant difference between the groups L and G (P<0.05).The MCP-1 mRNA expression in lung tissues was higher in the control group (P<0.01), and there was also a significant difference between the groups L and G (P<0.01).Conclu-sions Diverse types of particulate matters induce different pathological changes in the lungs, therefore the mechanism may also be different in the inflammatory responses.It means that the lung injuries caused by fine particles of mixed composition may have complex mechanisms.