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 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.

To observe the effects of a Chinese compound herbal medicine for reinforcing qi, activating blood circulation, expelling phlegm and dredging collaterals in treatment of lower limb arteriosclerosis.

The clinical features of 172 female and 1 067 male residents with gout in the coastal regions of Shandong province were analyzed.The results showed that as compared with male patients the onset of gout occurred at an older age in the female patients,and there were less obesity and tophi as well as less alcohol consumption and medication needed in the female patients.However,comorbidity with diabetes mellitus and coronary heart disease,as well as involvement of upper limb and other metatarsophalangeal joints were more prevalent in the female patients.

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 prepare a three-pulse drug release system of nimodipine and study its drug release.METHODS:Solid dispersion technique and dry-coating method were respectively applied to prepare immediate-release mini-tablets and Pulsatile mini-tablets with lag-time of 4 h or 8 h.Then those mini-tablets were filled into capsule to obtain a three-pulse drug release system and subjected to in vitro dissolution test.DSC was employed to determine drug status in solid dispersion.RESULTS:Immediate-release mini-tablets were released more than 95% in 30 min.Pulsatile mini-tablets were released less than 10% in 4 h or 8 h of lag-time period.After lag-time period,pulsatile mini-tablets were released completely in 3 h.The whole pulsatile drug release system achieved three times of drug release at 5 min,4 h,8 h,respectively.Nimodipine kept amorphous form and were delivered into carrier evenly.CONCLUSION:A three-pulse drug release system of nimodipine has been prepared successfully.

Objective To discuss the risk factors for susceptibility of subcutaneous tophi with an aim to provide clinical evidence for the prevention and treatment of tophi.Methods A total of 5 321 cases of gout patients whose course of disease was less than 10 years were selected and divided into two groups according to whether a subcutaneous tophus was present. The clinical information was collected and relevant biochemical indices were detected.Results There were significant differences in the ratios of regular exercise, involvement of upper limb joints, combining renal insufficiency, kidney stone and coronary heart disease between the tophus group and the non-tophus group(P<0.05 or P<0.01). There were significant statistical differences in age, disease duration, body mass index, systolic blood pressure, diastolic blood pressure, serum uric acid, serum creatinine, creatinine clearance rate, the number of joints involved between the two groups[(51.80±14.58 vs 48.45±14.45)year,(5.61±3.20 vs 3.28±3.06)year,(26.50±3.39 vs 27.05±3.45)kg/m2,(138±20 vs 133±18)mmHg, 1 mmHg=0.133 kPa,(89±13 vs 87±12)mmHg,(486.66±121.96 vs 460.02±119.10) μmol/L,(112.11±62.97 vs 104.43±46.28) μmol/L, (86.46±37.99 vs 93.00±40.11 )ml/min,(2.89±1.86 vs 1.76±1.22) joints, all P<0.01]. There were no statistical differences in family history, gender, history of smoking and drinking, the means of waistline, fasting blood glucose, triglyceride, and total cholesterol between the two groups(all P>0.05). Further logistic regression analysis indicated that disease duration, a large number of joints involved, the involvement of upper limb joints, combining kidney stones, higher serum uric acid level, and higher diastolic pressure were risk factors of the early onset of subcutaneous tophi, while regular exercise and body mass index were protective factors.Conclusion Patients with long duration, high serum level of uric acid, a large number of joints involved, upper limb joint involved, kidney stones, and hypertension were more likely to develop subcutaneous tophi. These risk factors should be intervened actively in clinic.