Objective To construct yeast eukaryotic expression vector carrying human endostatin (ES) cDNA. Methods The functional fragment of endostatin gene in human hepatic tissue was amplified by using RT-PCR technology, and cloned into yeast pPIC9 expression vector. The positive clone was sequenced by using automatized sequencer. Results The endostatin cDNA was successfully cloned. The positive ES clone gene in pPIC9 expression vector was sieved, and its coding sequence was identified to be as same as the previously reported sequence. Conclusion The successful construction of ES gene in pPIC9 expression vector using molecular biological method maybe helpful for the high expression of ES protein, which may lay the foundation for the treatment of malignant tumor through anti-angiogenesis appoach.

Objective To evaluate the effect of bone marrow stem cells (BMSCs) transplantation in the treatment of hepatolenticular degeneration of liver fibrosis. Methods Sixty cases with confirmed hepatolenticular degeneration of liver fibrosis were randomly divided into 3 groups: penicillamine group [40 mg/(kg·d)], BMSCs group and BMSCs + penicillamine group. Autologous BMSCs (2 mL) were injected into vein with normal saline (100 mL). Liver tissue pathology biopsy was inspected and the changes in HA, PCⅢ, LN, CⅣ, TIMP-1 and MMP-1 were observed at 0, 4th, 8th and 12th week during the therapeutic process. Results The level of serum fibrotic markers were reduced in every group , while the changes in BMSCs + penicillamine group were especially obvious (P < 0.05). Conclusion Penicillamine combined with BMSCs was effective in the therapy of hepatolenticular degeneration of liver fibrosis.

Objective To improve clinicians' understanding of tuberculosis as the adverse event of the tumor necrosis factor alpha (TNF-α) antagonists therapy. Methods Two rheumatoid arthritis (RA) patients were reported to develop lymphoid tuberculosis and pulmonary tuberculoma after TNF-α antagonists therapy. The relevant literature were reviewed. Results The first ease was a 62 year-old female RA patient. Her skin PPD reaction was negative and chest X-ray was normal before the therapy. Following 4 doses of infliximab injection (each dose of 3 mg/kg) which were completed within 3 months, the patient developed fight supraclavicular lymphoids tuberculosis in the 5th month after the last treatment. The patient completely recovered after lymphoids excision and antituberculosis therapy with 4 anti-tuberculosis medications combination. The second case was a 44 year-old female RA patient. She did not take the skin PPD test and chest X-ray before the therapy. The patient developed fever and chest distress after etanercept therapy (25 mg hypodermic twice per week ) for 1.5 months. The chest X-ray showed a shadow in the median lobe of the right lung, which had gradually developed to a pulmonary tuberculoma. The patient's physical condition improved after the tubereuloma resection. It has been reported in recent years that ① TNF-α antagonists therapy could increase the incidence of tuberculosis, ② the incidence at which infliximab associated tuberculosis was higher than etanercept, ③ the majority of the patient having tuberculosis were old people, and ④ the incidence of extrapulmonary tuberculosis and the disseminated tuberculosis were higher than regular tuberculosis. Conclusion TNF-α antagonists may decrease the host defense ability against mycobacterium tuberculosis and increase the incidence of tuberculosis. The pre-treatment tuberculosis screening, as well as tuberculosis monitoring during and after treatment is mandatory.

Objective To explore the influence of Hedysari radix concentrations fluid (HRCF) on learning and memory and its effect on cerebral monoamine neurotransmitters in senescence accelerated mice. Methods The senescence accelerated mice were randomly divided into the senescence accelerated mouse/resistance 1 (SAMR1) control group, senescence accelerated mouse/prone-8 (SAMP8) model group, HRCF treatment group and aricept treatment group. After 3 months of intragastrical treatment, the learning and memory ability changes and content of norepinephrine, dopamine, 5-hydroxytryptamine and 5-hydroxyindoleacetic acid were detected respectively using Morris water maze and high performance liquid chromatography. Results In the model group, the hidden platform test indicated that the latency to find the hidden platform was remarkably prolonged, and the content of norepinephrine, dopamine, 5-hydroxytryptamine as well as 5-hydroxyindoleacetic acid were significantly reduced (P<0.01). Compared with the model group, the latency of hidden platform test was remarkably shortened (P<0.05) and the content of norepinephrine, dopamine, 5-hydroxytryptamine as well as 5-hydroxyindoleacetic acid were obviously increased in HRCF treatment group (P<0.01). Conclusion HRCF could improve learning and memory ability of SAMP8 and the influence may related to increasing the content of cerebral monoamine neurotransmitters.

Objective:To evaluate the influence of bone marrow stem cells ( BMSCs ) in the serological indicators of hepatolenticular degeneration combined liver fibrosis.Methods:60 cases were randomly divided into 3 groups: penicillamine group, BMSCs group and BMSCs+penicillamine group.BMSCs(2 ml)were injected into vein with normal saline(100 ml) every 10 days ( 3 times for a period of treatment).Liver tissue pathology biopsy was inspected and TBIL, ALT, ALB, CHE, PDGF-BB, TGF-β1, IL-6 and TNF-αwere detected at 0,4 ,8 and 12 weeks.Results: The level of serological indicators about liver functions were reduced in every group, while the changes in BMSCs+penicillamine group were especially obvious ( P<0.05 ).Conclusion: Penicillamine combined with BMSCs was effective in the improvement of liver functions of hepatolenticular degeneration combined liver fibrosis.

OBJECTIVE:To study the analgesic effect of total flavonoids of propolis(TFP)and its mechanism.METH?ODS:The analgesic effect test is performed by formaldehyde method,hot-plate method,warm bath method and writhing method.The mice were randomly divided into normal saline(NS)control group,morphine positive control group and different dosage of TFP group,each pain index under each experiment condition was determined after intragastric administration of each group of mice.RESULTS:Compared with the NS control group,TFP group showed a decreased scoring in pain reaction(15min later),reduced writhing response(50,100mg/kg),prolonged latent phases of foot licking(50,100mg/kg)and tail-shrinking(50mg/kg),significant decrease of the content of malonaldehyde,prostaglandin-2and NO in mice'blood serum and brain tissue.CONCLUSION:TFP have notable analgesic effect,the mechanism may be related to its function on inhibiting prostaglandin-2and lipid peroxidation and reducing the release of NO releasing of brain tissue.

Objective To investigate the improving effect of Hedysari radix polysaccharide on the behavior and its influence on cerebral central neurotransmitters in senescence accelerated mouse prone 8 (SAMP 8).Methods The senescence accelerated mice 8 were randomly divided into a model group,a Hedysari radix polysaccharide (HRP) group and a aricept group.SAMRl were served as the control group.After intragastrical treatment for 3 months,the behavior changes and the levels of norepinephrine (NE),dopamine (DA),5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were observed respectively by applying Morris water maze and high performance liquid chromatography (HPLC).Results In the SAMP8 model group,the hidden platform test showed that the latency to find the hidden platform was remarkably prolonged,the latency range from third day to fifth day was (55.31 ±9.62) s,(58.67± 10.89) s,(58.45± 10.53) s,respectively.The level ofNE、DA、5-HT and 5-HIAA was (4.97± 1.74) mmol/L,(1.63±0.57)mmol/L,(4.29±0.94)mmol/L,(2.91±0.85)mmol/L respectively,significantly reduced (P＜0.01).Compared with model group,the latency of hidden platform test was (44.73 ± 9.83) s,(40.53±8.37)s,(44.75±9.16)s respectively,which were remarkably shortened.The expression level ofNE,DA,5-HT and 5-HIAA in the HRP treatment group was (7.58± 1.62) mmol/L,(3.25 ±0.70) mmol/L,(7.04±0.95) mmol/L,(4.62±0.79) mmol/L respectively,which obviously increased (P＜ 0.05).Conclusion HRP could improve learning and memory ability of SAMP8 and the effect may related to up-regulate the expression level of cerebral Central neurotransmitters.

Objective To introduce a new extracapsular approach for thyroidectomy in patients with surgical thyroid disease. Method Clinical data of 104 cases undergoing this procedure were retrospectively analyzed. Results From May 1990 to May 2000, 104 patients underwent either total or subtotal thyroidectomy by extracapsular approach. One patient suffered from postoperative temporary recurrent laryngeal nerve palsy. No morbidity such as postoperative thyroid crisis, hemorrhage leading to airway compression, permanent hypoparathyroidism nor mortality developed. Conclusion The total extracapsular approach for thyroidectomy is safe, less of complication and effective.

Objective To compare the effect of intra-articular injection of recombinant human tumor necrosis factor-Fc (rhTNFR:Fc) in the treatment of knee arthritis in spondyloarthritis (SPA) with that in rheumatoid arthritis (RA).Methods The subjects included in this study were SpA and RA patients with knee arthritis without deformity,moderate or severe bone erosion and obvious joint space narrowing in radiography,who had taken at least 6-week therapy with routine dosage of disease modifying anti-rheumatic drugs (DMARDs) before the study.All subjects received one dose of 25 mg rhTNFR:Fc injected to the target knee joints after synovial fluid being drawn away before injection.They were followed up for four weeks after injection.The primary end-point was the 4-week change in the moditied hospital for special surgery (HSS) knee score for the target knee.The secondary end-point were the 4-week change in patients assessment of the knee,and investigators assessment of knee,pain VAS of the knee joint when walking or standing,the range of knee inflexion,circumference of the knee cross section,synovium thickness detected by ultrasound in the part of the thickest synovium in the suprapatellar bursa.Paired t test,independed t test and Wilcoxon test were used for statistical analysis.Results Twenty-seven SpA patients and fifteen RA patients were included in the trial.The modified HSS knee score for the SpA group was 66±14 at baseline,86±11 (P＜0.05) at follow-up,and for the RA group,the score was 64±13 at baseline,80±9 (P＜0.05) at follow-up.24.2％ (16.5％-41.9％) improve-ment on the modified HSS knee score was achieved in the SpA group,while 22.2％ (15.3％-37.7％) improvement on the modified HSS knee score was achieved in the RA group (P＞0.05).31.8％ (9.3％-57.3％) improvement on the synovium thickness was achieved in the SpA group,while 1.5％ (-19.3％-25.5％) improvement on the synovium thickness was achieved in the RA group (P＜0.05).Adverse events were observed in six patients in SpA group and two patients in RA group.No serious adverse events had been observed.Conclusion Single intra-articular rhTNFR:Fc injection is an effective and safe therapeutic option for knee arthritis in both SpA and RA patients.This treatment may relieve knee synovitis in SpA patients more effectively than in RA patients.

Objective To study the anti-fibrotic function and mechanism of peroxisome proliferator activated receptorγ(PPARγ) in connective tissue disease-interstitial lung disease (CTD-ILD).Methods The expression of PPARγin lungs was analyzed in 37 cases with CTD-ILD and 20 control cases by immunohistochemistry.Changes in α-SMA levels were analyzed by Western blotting,and acetylation of Smad3 and Smad3 or PPARγ combined with P300 were analyzed by IP-WB.The data was analyzed by one-way ANOVA,t test or Mann-Whitney test.Results PPARγ' expression in the lung of CTD-ILD was lower than the controls [0.92％(1.44％),3.50％(1.94)％,respectively; Z=-8.924,P＜0.01].Different concentration of PPARγ (0,1,5,10,20,40 pmol/L) ligandinhibited the marked elevation of the protein α-SMA induced by TGF-β1 in a concentration-dependent manner (0.918 ±0.062,0.852±0.042,0.725 ±0.057,0.678 ±0.042,0.418 ±0.022,0.456±0.029; P＜0.05 or P＜0.01).However,this response was blocked by a selective antagonist PPARγ signaling GW9662 (0.946±0.087 vs 0.538±0.120,P＜0.01).Acetylation of Smad3 expression was increased when TGF-β1 was putted into lung fibroblasts after 60,90 and 180 min (0.565±0.047,1.127±0.101,0.873±0.022,0.614±0.407; all P＜0.05).The combination of Smad3 with P300 was also increased (1.46±0.12,0.98±0.09; P＜0.05),compared with the controls.But the ligand of PPARγ could block this effect (0.62±0.10,1.46±0.12; P＜0.05).Meanwhile,the combination of PPARγ and P300 was increased (0.94±0.05,0.76±0.22; P＜0.05).Conclusion PPARγ may play a physiologic role in the regulation of anti-fibrosis response.Its function may be realized by its competition with Smad3 combined with P300.