Objective　To approach the relationship betwe en glycosaminoglycan metabolism in cartilages and pathogenesis of KBD.Methods　Rhesus monkey was fed with grains and water from KBD endemic area for 18 months to produce the animal model with KBD . The glyc osaminoglycans in the monkey cartilage were extracted by the improved Dish method of Bitter. Purified glycosaminoglycans were digested with chondroitinaseABC, and the enzymatic digests were analyzed by HPLC. Results　Comparing with those of the control, the glycos aminoglycans in the head of femur, tibia plateau and costal cartilage from the Rhesus monkey fed with grains and water from KBD endemic area were undersulfated . Decreased unsaturated 4-sulphated disaccharide (△Di-4S) from the glycosa minoglycans in the head of femur and tibia plateau and decreased unsaturated 6- sulphated disaccharide (△Di-6S) from the glycosaminoglycans in the costal cart ilage were discovered.Conclusions　Detrimental factors in grains and water from KBD endemic area cause undersulfate of the cartilages glycosaminoglycans from Rhesus monkey. The glycosaminoglycans changes have a direct bearing on the patho logical alterations in morphology on the cartilages from the animal model with K BD.

Objective: To investigate the effect of combining acupuncture and auricular point sticking on heart rate variability (HRV) in patients with post-stroke depression (PSD). Methods: A total of 80 cases with PSD were randomized into a treatment group and a control group. The control group was intervened by oral administration of paroxetine hydrochloride, whereas the treatment group received acupuncture plus auricular point sticking base on the same oral administration. The Hamilton depression rating scale (HAMD) and HRV were measured before and after treatment in both groups. Results: The individual and global scores of HAMD significantly dropped after 8 weeks of treatment in both groups (all P<0.05). In the treatment group, anxiety/somatization factor, sleep disturbance, hopelessness factor, cognition factor and global score were significantly different from those in the control group (all P<0.05). The 24 h standard deviation of all normal-to-normal R-R interval (SDNN), standard deviation of 5-minute average of normal R-R intervals (SDANN), root mean square of successive differences (RMSSD), percent of differences between adjacent normal R-R intervals >50 ms (PNN50) and high frequency (HF) were increased while low frequency (LF) and LF/HF decreased significantly after 8 weeks of treatment in both groups (P<0.05). All items in the treatment group were significantly different from those in the control group (all P<0.05). Conclusion: Combining acupuncture and auricular point sticking can enhance the conventional medical treatment for HRV in patients with PSD.

Rutaecarpine (Rut) is a type of indole quinazoline alkaloid exracted from Ruticarpum. Studies showed that Rut has a wide range of pharmacological effects, such as anti-hypertension, anticancer, anti-inflammation, anti-thrombus formation. Currently, many scholars are committed to developing it into a new antihypertensive and anti-inflammatory drug with all new mechanisms. But studies found that Rut is a highly fat-soluble drug with low water and oil solubility. Its high insolubility is the main obstacle in its oral absorption and application, which greatly reduced its bioavailability. Therefore, hydroxypropyl-beta-cyclodextrin (HP-beta-CD) was used as the inclusion material to prepare Rut-HP-beta-CD inclusion complex in this experiment, in order to increase its water solubility and bioavailability. In this experiment, the inclusion complex was prepared by the stirring-freeze-dry method. The preparation process was optimized by the orthogonal test, with the inclusion rate as the index, and molar ratio between host and guest molecules, inclusion temperature, time and stirring speed as the impacting factors. Moreover, the inclusion complex was verified by detecting the apparent solubility, thin layer chromatography, microscopic identification, melting point detection and dissolution study. The results showed that under the conditions of the molar ratio between Rut and HP-beta-CD of 1: 1, temperature at 60 degrees C, inclusion time of 4h and stirring speed at 600 r x min(-1), the inclusion rate of Rut-HP-beta-CD reached 91.04%. Therefore, the preparation process of Rut-HP-beta-CD inclusion under the optimum conditions is simple and feasible, with a highest inclusion rate and reproducibility, and could significantly improve Rut's solubility and bioavailability, and provide a reliable experimental basis for its clinical application.
2-Hydroxypropyl-beta-cyclodextrin ; Alkaloids ; chemistry ; Chemistry, Pharmaceutical ; methods ; Drug Carriers ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Rutaceae ; chemistry ; Solubility ; beta-Cyclodextrins ; chemistry

Microbial lipase,one of important industrial biocatalysts,has been used widely in many industrial and agricultural fields.It is always the research focus to screen,mine and develop the microbial lipases with novel catalytic activity and high stability.This paper introduces briefly the pathways and methods to mine novel microbial lipase resources from six aspects,including extremophile,metagenome,genome database,protein engineering,immobilization,chemical modification,etc.

Microbial lipases are important industrial biocatalysts with the character of stereoselectivity,site selectivity and high catalytic activity with few side effects.They have been used widely in many industrial and agricultural fields.The technology of protein engineering has been successfully applied to improve the activity and stability of microbial lipases,which will raise the competitive capacity of microbial lipase preparations and enlarge theirs application fields.The strategies,the problems and the prospects of protein engineering technology which have been applied to modify the microbial lipases was surveied.

OBJECTIVE To investigate the anti- tremor effect and mechanism of baicalein on oxotremorine- induced muscle tremor in mice. METHODS The acute model of muscular tremor was induced by intraperitoneal injection of oxotremorine, and the latency, duration and frequency of muscle tremor in mice were measured immediately; the saliva of mice was measured to reflect the correlation between tremor and peripheral nerve function; the aim of this study was to determine the content of MDA and the activity of GSH-PX, and to investigate the anti-oxidation of mice with tremor model. The activity of acetylcholinesterase (AchE) and acetylcholine transferase (ChAT) can indirectly reflect the level of acetylcholine in the brain. The level of monoamine neurotransmitters in brain tissue was determined by high performance liquid chromatography (HPLC-ECD). RESULTS The animals in the model group appeared obvious tremoring, salivating and erecting and other symptoms. Compared to the model group, there was no obvious inhibitory effect on the administration of each dose. After 7, 14, 21 and 28 d of continuous administration, the latency, duration and tremor frequency of tremor mice were significantly shortened, the levels of acetylcholine were significantly decreased, the changes of DOPAC and DA neurotransmitters in the brain of model group were recovered, regulate the dynamic balance of acetylcholine and dopamine in the brain. CONCLUSION Long- term administration can improve the tremor behavior of mice, the mechanismmay be related to the regulation of neurotransmittersin brain.

Objective To investigate the effects and mechanisms of 1,25-Dihydroxyvitamin D3 [1,25(OH)2 D3] on hyperoxia-induced lung injury of neonatal rats.Methods Neonatal Sprague-Dawley rats were randomly divided into air group,hyperoxia group and 1,25(OH)2D3 group within 12 hours after birth,eight in each group.Rats in air group were exposed to air,while those in hyperoxia and 1,25 (OH) 2 D3 group were exposed to hyperoxia (≥85 ％ oxygen concentration).Rats in 1,25(OH)2D3 group were injected with 1,25(OH)2D3 0.5 μg/(kg · d) intraperitoneally once a dayfor seven days,meanwhile the rats in the other two groups received 0.9 ％ saline in the same way.All rats were sacrificed on day 7.Lung tissue sections were HE stained in order to assess lung histological changes and lung radical alveolar count (RAC).The levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β and IL-6 mRNA were measured by real time-polymerase chain reaction.Analysis of variance and LSD test were applied for statistics.Results Compared with the air group,the weight of rats in the hyperoxia group was significantly lower on day 7 [(8.48±1.34) g vs (12.51±0.47) g,t=8.05,P＜0.05],while the weight of rats in 1,25(OH)2D3 group [(10.29±1.00) g] was higher than that in the hyperoxia group (t=3.61,P＜0.05).Lung tissue structure was normal in the air group.In the hyperoxia group,inflammatory exudation was observed in pulmonary interstitial,the alveolar size was uneven,and the RAC was lower than that in the air group (5.6±0.1 vs 6.8±0.2,t=21.45,P＜0.05).The RAC in 1,25(OH)2D3 group (6.2±0.1) was significantly increased compared with that in the hyperoxia group (t=11.76,P＜0.05),but still lower than that in the air group (t=9.69,P＜0.05).The expressions of TNF-α,IL-1β and IL-6 mRNA in hyperoxia group (0.0348±0.0006,0.0269±0.0003 and 0.0368 ± 0.0006) were higher than those in the air group (0.0111±0.0007,0.0040±0.0003 and 0.0162 ±0.0007,t=56.54,111.12 and 49.26,P＜0.05,respectively).The expressions of TNF-α,IL-1β and IL-6 mRNA in the 1,25 (OH)2D3 group (0.0203±0.0009,0.0141±0.0004 and 0.0251±0.0009) were lower than those in the hyperoxia group (t=34.44,61.93 and 27.99,P＜0.05,respectively),but higher than those in the air group (t=22.10,49.19 and 21.27,P＜0.05,respectively).Conclusions 1,25(OH)2D3 could attenuate hyperoxia-induced lung injury by inhibiting the expression of inflammatory cytokines.

Purpose To study the malignant transformation after treating rat oval cell line ( WB-F344 ) with chemical carcinogen N-methyl-N′-nitro-N-nitrosoguanidine ( MNNG) . Methods WB-F344 cells were cultured with MNNG for severe times. The biological characteristics of induced cells were detected through the following methods:to check proliferation activity by flow cytometry analysis, to examine malignant transformation degree of induced cells by soft agar assay and tumor formation in NOD/SCID mice, and to investi-gate the transcriptional and protein levels of hepatocellular carcinoma marker GGT, GST-P by real time-PCR. Results Oval cells in-duced by MNNG showed changes in biological characteristics and malignant molecular markers. Conclusion Hepatic oval cells model is successfully established, which can be confirmed by tumor formation in NOD/SCID mice.

ABSTRACT:Objective To observe the effects of late Na current (INa‐L ) and rapidly activating delayed rectifier K current (IKr ) on ventricular heterogeneity and frequency dependency by using high resolution voltage sensitive optical mapping technology .Methods The model of 12 isolated hearts was constructed in rabbits . Voltage sensitive dye Di‐4‐ANEPPS were perfused into the isolated hearts by Langendorff method .LED source with the wave length of 532 nm was used to record APD80 and APD50 of the left and right ventricles .Experimental groups were divided into 3 groups by perfusion drugs dofetillide (30 nmol/L) ,dofetillide+ATX‐Ⅱ(1 nmol/L) ,and dofetillide +ATX‐Ⅱ +mexiletine (10μmol/L) .The subjects were intervened by the above drugs in order ,and they were self‐compared before dosing .After each drug administration ,the hearts were stimulated respectively with the BCL of 2 000 ms ,1 000 ms ,500 ms ,and 300 ms .Then we observed the changes of APD80 and APD50 in the left and right ventricles before and after the interventions .Results ① In the control group ,APD80 and APD50 of the right ventricle were longer than those of the left ventricle in response to different stimulation , and the differences increased with the decrease of stimulating frequency .② When BCL was 1000 ms ,APD80 and APD50 of the left and right ventricles were prolonged respectively after administration of dofetillide , but the differences in APD80 and APD50 were insignificant between the left and right ventricles (P>0 .05) .ΔAPD80 of the two ventricles increased significantly with the decrease of stimulating frequency . ③ After administration of ATX‐Ⅱ , when BCL was 1000 ms ,APD80 and APD50 of the left and right ventricles increased significantly compared with those in the control group and dofetillide intervention group (P<0 .05) .And the increase of APD in the left ventricle was greater than that of the right ventricle .ΔAPD80 of the two ventricles increased significantly with the decrease of stimulating frequency .④ After administration of mexiletine ,when BCL was 1000 ms ,APD80 and APD50 of the left and right ventricles reduced significantly compared with those of the primary state (P<0 .05) .APD80 and APD50 of the left and right ventricles reduced significantly compared with those of the control group (P< 0 .05) and ATX‐Ⅱ group (P>0 .05) .The increase of ΔAPD80 of the two ventricles became milder when the stimulating frequency decreased . Conclusion ① IKr blocked by dofetillide did not affect the heterogeneity between the two ventricles , which showed reverse‐frequency dependence . ② In the context of blocking IKr , ATX‐Ⅱ increased the heterogeneity between the left and right ventricles and enhanced the reverse‐frequency dependence .In contrast ,mexiletine ,the blocker of INa‐L ,decreased the heterogeneity between the two ventricles and reverse‐frequency dependence .

Objective：To study the clinical and pathologic features of patients with lupus nephritis (LN) whose myeloperoxidase anti-neutrophil cytoplasmic antibody (MPO-ANCA) were positive. Methods：The clinical and pathological features were analyzed in 18 patients with LN whose MPO-ANCA were positive. And the data of patients with different clinical outcomes were compared. Results：（1）The hematological abnormalities, hypertension and serositis in these patients were more common than general ones with LN. （2）Proteinuria and hematuria were common, the morbidities of gross hematuria and renal failure in these patients were higher than general ones with LN.（3）Various autoantibodies were positive in these patients.（4）Segmental necrosis crescentic nephritis accompanied by density of immunocomplex in glomeruli and vasculitis in intestitium were common.（5）The morbidity of ESRF and mortality of these patients were similar to general ones with LN. The morbidity of tubular atrophy in those with poor prognosis was significantly higher than those survived. Conclusion：The patients with LN whose MPO-ANCA are positive have some difference from those with negative MPO-ANCA, but positive MPO-ANCA is not directly related to the prognosis.