Rcsult of field cultivation test by protoplastic fusion technique for biological cngln ering breeding of Poria cocos in Huoshan county, Anhui Province was reported for the first time. A fusion strain F1 was obtained from parents P1, P6578 by separation, fusion, marker of hypha protoplast and identification of recombinat. It was cultivated simultaneously with P1, P5778 for comparison. Results showed that the average cellal outputs were 1.73, 0.5 and 2.20kg respectively.As the output of strain F1 is intermediate between its parent P1 and P578, the result is considered to be unsatisfactory. The reason for such result was discussed.

Objective: To evaluate the relationship between the levels of serum uric acid (SUA) and prevalence of coronary artery disease (CAD) by Meta analysis. Methods: We searched the databases of Pub Med, Elsevier and Web of Science for internationally published cohort study for the relationship between SUA levels and CAD prevalence and conducted a general analysisby using Stata software. Results: A total of 11 cohort study including 463,918 subjects were enrolled in this study. For both male and female genders, increase SUA level was the risk factor for CAD occurrence (RR=1.11, 95% CI 1.00-1.24) and (RR=1.24, 95% CI 1.15-1.34). Dose-response Meta-analysis indicated that by 1 mg/dl SUA elevation, the risk of CAD occurrence would increase 4.8% in male and 12.4% in female, the risk in female gender was higher than male. Conclusion: SUA level has been closely related to CAD prevalence.

OBJECTIVE:To study the effect of tanshinone on inflammatory response in air-pouch model mice with artificial joint aseptic loosening. METHODS:Mice were randomly divided into blank control group(normal saline),titanium particle group (normal saline),tanshinone low-dose,medium-dose,high-dose groups(50,100,200 mg/kg),10 in each group. Air-pouch mod-els were induced. Except that mice were injected 0.5 mL normal saline into air-pouch in blank control group,other groups were in-jected 0.5 mL Titanium particle suspension(10 mg/mL)into air-pouch,continuously administrated medicines by 0.1 mL/10 g after 24 h,ig,for 14 d. After 24 h of last administration,the air-pouch was collected,air-pouch inflammation was observed by eyes and by microscopy after hematoxylin-eosin staining,and inflammatory cell density was calculated. Real-time quantitative poly-merase chain reaction method was conducted to detect the tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β)mRNA expres-sion;enzyme-linked immunosorbent method was used to detect the TNF-α,IL-1β protein expression. RESULTS:Compared with blank control group,air-pouch swelling was obvious in titanium particle group,much exudation and neovascular were observed,in-flammatory response was severe,inflammatory cell density was increased significantly(P<0.05);TNF-α,IL-1β mRNA and pro-tein expression were obviously enhanced(P<0.05). Compared with titanium particle group,air-pouch swelling was relieved in tan-shinone doses groups,exudation and neovascular were decreased,inflammatory response was relieved,inflammatory cell density was decreased significantly(P<0.05);TNF-α,IL-1β mRNA and protein expression were obviously decreased(P<0.05),with a dose-dependent manner. CONCLUSIONS:Tanshinone can effectively inhibit the aseptic inflammatory response in air-pouch model mice with artificial joint aseptic loosening.

Objective To study the effect of the extract of total flavonoids of Chrysanthemum indicum(TFC) on adjuvant arthritis(AA) synoviocytes.Methods Totally 0.1ml of the complete Freund's adjuvant was subcutaneously injected into the right hind feet pads of 20 SD rats.24 days after immunity synoviocytes in the knee joint were treated with TFC.Cell morphology was examined with electron microscopy.Protein level of caspase-3 cleaved fragments was analyzed by Western blotting.The annexin V stain assay was applied to explore the effect of caspase-3 inhibitor on the amelioration in synovial cells apoptosis of AA rats.Results Typical morphology and biochemical feature of apoptosis in synovial cells of AA rats were observed with TFC.The protein level of caspase-3 cleaved fragments increased obviously and was related with the concentration of TFC in synovial cells of AA rats.The apoptotic cells positively stained with annexin V were markedly reduced by caspase-3 inhibitor.Conclusion TFC can induce apoptosis in AA rats synoviocytes,which may achieve therapeutical effects in AA.The activation of caspase-3 may be one of the main causations.

Objective To analyze the clinical characteristics of familial and sporadic gout patients to provide information for the classification,individual treatment and prognosis of gout.Methods The clinical and biochemical characteristies of 431 patients with familial gout and 1899 patients with sporadic gout were compared and analyzed.T test and X2 test were used for statistical analysis.Results The age at onset [(47±13) years ] and the serum uric acid level [ (472±125) μmol/L] of the familial gout patients in the acute phase were significantly lower than those of the sporadic gout patients (P＜0.05).The percentage of patients whose attack were induced by purine-rich food (67.7％ vs 88.2％),drinking (31.3％ vs 44.5％) and the first metatarsophalangeal joint involvement (69.1％ vs 77.4％ ) were significantly lower in the familial gout patients than those in the sporadic group of patients.The percentage of female was lower in the familial gout (9.7％) than in the sporadic patients(6.6％,P＜0.05).The percentage of patients with ankle joint (18.1％ vs 11.3％) and no obvious predisposing causes (25.2％ vs 2.5％ ) were higher in the familial gout patients than in the sporadic patients (P＜0.05).The percentage of patients with complicated lipid metabolism disorders was significantly higher in the familial gout patients than in the sporadic patients (P＜0.05).Conclusion The familial gout patients in Shandong coastal regions are early in disease onset,with lower serum uric acid level and more frequent in women.Detailed family history should be collected,and early prevention and appropriate treatment should be emphasized.

Objective To Explore the significance of successful exposing and recognizing Zuckerkandl's tuhercle(ZT)during thyroidectomy.Methods Three hundred and seventy patients(501 sides) underwent lobectomy or total thyroidectomy from January 2009 to June 2011 were included in this study.The ZT was assessed in terms of its presence or absence,size and anatomical association with the recurrent laryngeal nerve(RLN)and superior parathyroid(SP).Results ZTs were found in 412 of 501 sides ( 82.2％ ),among which 368(89.3％ ) ZTs were located in the middle third of the lateral lobe of the thyroid gland.ZTs passed over the RLN in 379 of 412 sides(92.0％ ).When the ZTs were located in the middle or lower third of the lateral lobe of the thyroid gland,the SPs were all located in the cranial portion of ZT.The SP was adhered to the ZT in 80.1％ of the cases.RLN damage rate was 0.40％,and no SP damage occurred.Conclusion Exposing and recognizing Zuckerkandl's tubercle during thyroidectomy is of important clinical significance,which helps to identify and protect RLN and SP,so as to reduce surgical complications.

This study is to investigate effects of genistein on rat femoral bone metabolic in vitro. Rat femoral tissues was isolated and randomly divided into two groups including control group and genistein (1 x 10(-5) mol x(-1)) group. Determinations of alkaline phosphatase (ALP) activity, calcium content and osteoprotegerin (OPG), type I-collagen (Collagen-I), RANKL, Runx-2 and bone morphogenetic protein (BMP-2) mRNA expression were done by real-time PCR. The results showed that 1 x 10(-5) mol x L(-1) genistein could increase the activity of ALP and contents of Ca, regulate bone metabolism activity of OPG, RANKL, BMP-2, Collagen-I and Runx-2 mRNA expression level. Genistein can significantly modulate bone metabolism related gene expression level of rat femoral tissue in vitro, and can increase calcium content and the activity of ALP.

BACKGROUND:Studies have shown that sacral nerve-root stimulation based on anodes block technique can effectively reconstruct the bladder voiding function of the rabbits with spinal cord injury. But the corresponding technology of stimulating electrode has not been reported so far.
OBJECTIVE:To design and develop the stimulating electrodes matching with both rabbit sacral nerve roots and anodal blocking technique, to observe the ultrastructure and morphological change of rabbit sacral nerve roots which implanted in electrode stimulation for a long-term and to assess the safety of stimulating electrodes.
METHODS:Thirty New Zealand rabbits were included, 10 rabbits were randomly selected from them and sacrificed after anesthesia, and then cut the anterior roots of bilateral S 2 and S 3 immediately;after measuring the diameter under the light microscope, the sleeve type stimulation electrode matched with the diameter was made. The remaining 20 rabbits were randomly divided into control group and implantation group, with 10 rabbits in each group. In the implantation group, the stimulating electrodes were implanted into the forepart of S 2 and S 3 nerve roots after anesthesia, and then sacrificed after fed for half a year for col ecting the samples. Then ultrastructure change of sacral nerve roots with the implantation was observed.
RESULTS AND CONCLUSION:Structure of nerve cel s of sacral nerve roots remained in good condition under a light microscope after long-term implantation of the stimulating electrodes. No obvious degeneration of axons, no inflammatory infiltration and glial scar formation were observed. In the implantation group, myelins arranged closely without demyelination phenomenon, and there was no atrophy of neuronal nuclear, no nuclear sag, no increased nuclear decompression and heterochromatin in neurons under the light microscope. Immunohistochemical analysis showed, compared with the control group, there were no significant differences in the expressions of glial fibril ary acidic protein, Bax, Bcl-2 and Caspase-3 proteins of nerve roots in the implantation group. The stimulation electrode of rabbit sacral nerve root is developed successful y, that is, the implantation is simple and safe as it can be used for long-term implantation without histopathological changes and apoptosis.

Objective To study the impact of tumor necrosis factor-α (TNF-α) and its antagonist on the expressions of intestinal mucosa claudin-1,Zonula Occludens-1 (ZO-1) and myosin light chain kinase (MLCK) in rat models of acute liver failure.Methods Fifty four healthy male SpragueDawley (SD) rats were randomly divided into normal control group,model group and intervention group according to a random number table.Rats in normal control (n=6) group were intraperitoneally injected with 0.9％ saline (12 mL/kg).Rats in model group (n=24) and intervention group (n=24) were intraperitoneally injected with a full dose of D-galactosamine (D-GalN) at a dose of 1 200 mg/kg to establish model of acute liver failure,while rats in intervention group were intraperitoneally injected with TNF-α antagonists (rhTNFR∶Fc) at a dose of 12.5 mg/kg before 24 hours given D-GalN.At each time point of hour 8,24,48 and 72,six rats in both model group and the intervention group were sacrificed,respectively,while the normal control group were all anesthetized and sacrificed at 72 h.Models were repeated five times.Serum liver function was detected by biochemical method,and serum TNF-α level was detected by enzyme-linked immunosorbent assay (ELISA).Hematoxylin-eosin (HE) stained sections of liver and terminal ileum were examined under an optical microscope for pathological changes;and protein expression of the terminal ileum Claudin-1,ZO-1 protein and MLCK were determined by immunohistochemistry and Western blot.Means among groups were compared with t test.Results Acute liver failure was successfully induced in the D-GalN injected rats.In the model group,alanine aminotransferase (ALT) began to decline,total bilirubin continued to rise,and enzyme-jaundice separation developed at hour 72.But total bilirubin in intervention group at hour 72 was decreased.Light microscope showed that at hour 72,villus lodged at terminal ileum in the model group with part of villus tip failing off in the model group.Villus mucosa and submucosa interstitial were edema and infiltrated with numerous neutrophils.The terminal ileum kept integrate in the intervention group,and villus mucosa and submucosa were mild edema and only infiltrated with a small amount of neutrophil.Expressions of tumor necrosis necrosis factor (TNF)-α in rats of model group and intervention group were gradually increased and peaked at hour 24 ([239.83 ± 15.81] and [182.71± 17.08] ng/L,respectively),which were significantly higher than that of the control group ([24.19±3.57] ng/L,t=22.68and 15.73,respectively;both P＜0.01).Expression of serumTNF-α in the intervention group was significantly lower than that of model group (t=4.58,P＜0.01).Expressions of Claudin-1 and ZO-1 in model group decreased gradually at an early stage and reached the lowest level at hour 24 (0.355 ± 0.068 and 0.387 ± 0.091,respectively),which were both significantly lower than that of control group (1.640±0.188 and 1.015±0.150,respectively;t=12.87 and 7.14,respectively;both P＜0.01).In the intervention group,expressions of Claudin-1 and ZO-1 also decreased to the lowest level at hour 24 (1.051 ± 0.370 and 0.642 ± 0.082,respectivley),which were both significantly lower than that of control group (t =2.84 and 4.36,respectively;both P＜0.05),but significantly higher than model group with stastically difference (t =3.70 and 4.15,respectively;both P＜0.01).MLCK protein levels in the model and intervention group were gradually increased,which peaked at hour 24 (1.298±0.194 and 1.033 ± 0.073,respectively),significantly higher than the control group (0.460±0.069,t=8.16 and 11.44,both P＜0.01);and MLCK in the intervention group was lower than that in the model group with statistically difference (t=2.56,P＜0.05).Conclusions Expression of serum TNF-α in rat model of acute liver failure increases,which leads to decreased expression of Claudin-1 and ZO-1,and increased expression of MLCK,makes cell shrunk and cell gap increased.TNF-α antagonist could significantly reduce the inflammation and liver cell apoptosis,improve liver function by inhibiting MLCK expression and preventing decrease of Claudin-1 and ZO-1 proteins.

OBJECTIVE:To prepare Puerarin polymeric micelles and establish a method to determine its entrapment efficiency. METHODS:Puerarin polymeric micelles were prepared by film dispersion method. The polymeric micelles and free drug were sepa-rated by centrifugal-millipore filter filtration method. The entrapment efficiency of puerarin polymeric micelles was determined by HPLC. Diamonsil C18(2)column was used with 1% citric acid solution-methanol(65∶35)at the flow rate of 1 ml/min. The detec-tion wavelength was set at 250 nm,and column temperature was room temperature. RESULTS:The prepared polymeric micelles were spherical and spherical-like in shape with a mean particle size of 54.12 nm,polydispersity index of 0.122,Zeta potential of -13.60 mV;the linear range of puerarin was 2-10μg/ml(R2=0.999 4)with average recovery rate of 99.2%(RSD=0.9%,n=3). The re-covery rate of free drug was 95.3%(RSD=1.7%,n=3). The mean entrapment efficiency and drug-loading amount of puerarin were(35.5±2.12)% and(0.3±0.07)%,respectively(n=3). CONCLUSIONS:Film dispersion method is suitable for the prepara-tion of Puerarin polymeric micelles. Established method is convenient,accurate and reliable for the content and entrapment efficien-cy determination of Puerarin polymeric micelles.