EC-SOD inclusion bodies was refolded on column using metal (Ni) affinity chromatography, based on the metal-binding property of His-tag. The effect of protein amount, urea removal speed and temperature on refolding was observed. We compared the different efficiency purified with Ni-sepharose and Heprain-sepharose affinity chromatography, and studied the stability of the refolded proteins. The results indicate that the inclusion bodies can be renatured with Ni-sepharose affinity chromatography. The increase of the protein amount and urea removal rate , the lower of the renaturation efficiency. Higher temperature was benefit to protein renaturation. Both the Ni-sepharose and Heprain-sepharose affinity column can be used to purified the refolded proteins, but purified by Heprain-sepharose affinity column the protein had higher activity. The activity of renatured protein was stable in 10 ℃～50℃,when pH10 its stability was lower significantly. In denaturating solution the stability of renatured protein was low.

OBJECTIVETo explore the theory of "Fei and Dachang being interior-exteriorly related" and the pathogenesis of lung injury by observing changes of inflammatory cytokines and oxygen free radicals in ulcerative colitis (UC) rats.

METHODSTotally 50 healthy male Wistar rats were randomly divided into two groups, the normal control group and the model group, 25 rats in each group. The UC model was established by allergizing colon mucosa combined with TNBS-alcohol (50%) enema. Another 25 rats were recruited as the normal control group. At week 2 and 4 after modeling, the pathomorphological changes of the lung were observed. Furthermore, the contents of tumor necrosis factor alpha (TNF-alpha) and IL-1beta were determined by ELISA. The activities of superoxide dismutase (SOD) and malondialdehyde (MDA) were evaluated with colorimetry.

RESULTSCompared with the normal control group, the pathomorphology of the lung tissue in the model group appeared abnormal at week 2 and 4. Compared with the normal control group, levels of TNF-alpha, IL-1beta, and MDA in the lung tissue significantly increased in the model group (P < 0. 01) and the activities of SOD significantly decreased (P < 0.05, P < 0.01).

CONCLUSIONTNF-alpha, IL-1beta, SOD, and MDA might be common material bases for the large intestine involved in lung disease of UC patients, thus providing a modern scientific basis for the theory of Fei and Dachang being interior-exteriorly related.

Animals ; Colitis, Ulcerative ; diagnosis ; metabolism ; pathology ; Cytokines ; metabolism ; Interleukin-1beta ; metabolism ; Lung ; metabolism ; Male ; Malondialdehyde ; metabolism ; Medicine, Chinese Traditional ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Chemical constituents of Inula japonica were isolated and purified by repeated column chromatographies, over silica gel, and Toyopearl HW-40, and preparative HPLC. On the basis of spectral data analysis, including NMR and MS data, the structures of the isolates were elucidated and their anti-inflammatory activities were assayed. Fifteen compounds were isolated from the ethyl acetate extract of I. japonica, and their structures were elucidated as dihydrosyringenin (1), (3S, 5R, 6S, 7E)-5,6-epoxy-3-hydroxy-7-megastigmen-9-one (2), (6R, 7E) -9-hydroxy-4,7-megastigmadien-3-one (3), arnidiol (4), taraxasterol acetate (5), 8,9,10-trihydroxythymol (6), taxifolin (7), luteolin (8), napetin (9), eupatin (10), spinacetin (11), quercetin (12), p-hydroxycinnamic acid (13), caffeic acid (14), and caffeoyl acetate (15). Compounds 1, 2, 7, 13 and 15 were isolated from the genus Inula for the first time, and compounds 3, 4, 9-11 and 14 were isolated from this plant for the first time. The anti-inflammatory activity result showed that compounds 3, 6-12 and 14 exhibited inhibition effect against leukotriene C4 (LTC4) synthesis and degranulation definitely in c-Kit Ligand (KL) induced mast cells, and compound 8 and 12 also had the suppression effect against lipopolysacharide(LPS) induced nitric oxide (NO) activity in RAW264.7 macrophages. It is firstly reported that compounds 7 and 9-11 possessed potent inhibition activities against LTC4 generation and degranulation in mast cells.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacology ; Cell Line ; Inula ; chemistry ; Macrophages ; drug effects ; Mast Cells ; drug effects ; Mice ; Mice, Inbred BALB C ; Plant Extracts ; chemistry ; pharmacology

Objective To observe the effects of cultivation mode of autonomous learning ability in TCM formula on students’ learning capacity and comprehensive quality. Methods On the basis of inheriting the traditional teaching mode, classroom questioning, class discussion, problem-based learned teaching, simulated prescription exercises, autonomous teaching and training, the mid-term oral test, and autonomous proposition consisted of autonomous learning ability training mode diversification. In the teaching process, teachers' teaching was the primary, and was interspersed with cultivation mode of autonomous learning ability. At the end of the teaching activities, questionnaire survey method was adopted to evaluate the autonomous learning ability training mode in the teaching of TCM formula. Results 81.30%-97.74% students believed that this mode is helpful to improve their learning ability and comprehensive quality; 75.87%-98.49% students believed that it is necessary to carry out the mode in the teaching activities; 69.53% students believed that conducting mid-term examination in the form of oral teat is feasible. Conclusion The cultivation mode of autonomous learning ability for TCM formula can stimulate the learning enthusiasm of the students from various angles, raise interest in learning, arouse students’ learning enthusiasm and initiative, and further cultivate autonomous learning ability and clinical thinking ability, which can provide references for the teaching reform of TCM formula.

Postoperative pancreatic leakage remains a persistent problem after pancreaticoduodenectomy.For patients with a soft and nonfibrotic pancreas,double binding continuous hemstitch suture is an optimal method for anastomosis.From January 2011 to June 2012,92 cases of periampullary carcinoma with a soft pancreas underwent pancreaticoduodenectomy,and then a modified technique of pancreaticogastrostomy was performed with 2 continuous hemstitch sutures placed in the mucosal and seromuscular layers of the posterior gastric wall,respectively.The median time for pancreaticogastrostomy was 12 minutes,and only 15 patients had postoperative complications.Two patients developed pancreatic leakage(1 grade A and 1 grade B)postoperatively.Pancreaticogastrostomy with double binding continuous hemstitch sutures is a simple and safe reconstruction procedure for patients with a soft and fragile pancreas who received pancreaticoduodenectomy.

A fragment sized 400bp of White spot syndrome virus（WS SV，formerly de signated NOSV），recovered from recombinant plasmid pAFD, was labeled with Digox igenin as a probe to detect dynamic distribution of WSSV within 120h and 72h in crawfishes（Cambarus proclarkii） inoculated WSSV by oral taking and injecti on r espectively. Stomach epithelium, intestine epithelium, heart, gill, haemolymph, muscle, hepatopancreas, hypoderm, connective tissue and ovary of infected crawfi shes were examined for WSSV. In both groups, WSSV was first detected in heamoly mph at 12h p.i. and then disappeared. Again it was detected at 96h p.i. only in oral infection group and maintained till 120h p.i., but it didn't appear at 72h p.i. in injection group. WSSV in heart, muscle was detected at 36h p.i. in oral infection group and 24h p.i. in injection group respectively, and then increased generally. In addition, WSSV in intestine epithelium, connective tissue, ovary of oral infection group and intestine epithelium, hypoderm, ovary of injection g roup could also be detected. In dead crawfishes after 120h and 72h p.i. in two groups, WSSV could be detected in all the examined tissues and it demonstrated t hat systemic infection occurred in the animales. The tissue containing more amo unts of WSSV was hypoderm in oral infection group, while intestine epithelium, g ill, hypoderm, ovary in injection infection group. It deduced that WSSV first a ppears in haemolymph and then goes into heart, muscle and other tissues and prol iferates in them. Once again, WSSV is released into heamolymph resulting in syst emic infection till crawfishes' death.

Objective To observe the results and reality of transurethral resection of bladder tumor(TUR-Bt) with upside down loop and plsamakinetic in the prevention of obturator nerve reflex.Methods A total of 211 patients with lumbar anesthesia received TUR-Bt.Firstly,the tumor tower above bladder wall was cut with the traditional way.When intraoperative findings were obturator nerve reflex,or to the obturator nerve mapping area,operation should be changed to superimpulse plasma column electrode vaporization to cut off residual tumor tissue,then after flipping the loop,the residual tumour was resected with upside-down loop from the tumour side wall.Results In 211 cases,192 cases had a little obturator nerve reflex,but the movement of body was not obvious,which did not influence the operation.The serious complication,such as perforation of bladder and injury of nerve and vessel,did not occur.Conclusions The lateral resection of bladder side wall tumour with upside-down loop can effectively avoid strong obturator nerve reflex,which is safe,uncomplicated,and efficacious and it does not increase the additional outlay and hurt.

Objective To evaluate the results of NiTi shape memory alloy four-corner arthrodesis concentrator (NTMA-FCAC) for carpal collapse. Methods We reviewed retrospectively 13 patients who underwent scaphoid excision with four-corner arthrodesis using NTMA-FCAC for carpal collapse from August 2006 to June 2009. There were eight males and five females, with an average age of 38 years (range, 23-61years). The cause of carpal collapse was SNAC in 7 cases, perilunate dislocations in five and SLAC in one.The injury mechanisms included traffic accidents (5 cases), falling from a height (4 cases), crashes (3 cases)and sprain (1 case). Objective measurements included grip strength and range of the wrist. Radiographs were performed in all patients. A visual analogue scale (VAS) was used to assess wrist pain. The results were evaluated according to the Krimmer wrist scores. Results The mean follow-up time was 26.5 months (range,6-36 months). Clinical evaluation yielded the mean grip strength of (32.49±6.21) kg (80.8% of opposite side).The mean range of the wrist reached over 53% of the healthy side. Non-union and wound infection were not seen. The mean VAS scores had improved from 4.46±1.27 preoperatively to 1.31 ±0.95 postoperatively. The mean pain scores under stress had improved from 7.00±1.41 preoperatively to 2.62±1.26 postoperatively.There were remarkable differences between them. The mean Krimmer wrist score was 79. Conclusion Four-corner arthrodesis using NTMA-FCAC is an effective method for carpal collapse, preserving a majority of wrist function.

ObjectiveTo determine the impact of lupus flares on maternal and fetal outcomes in pregnant patients with systemic lupus erythematosus(SLE).MethodsData was obtained from 46 pregnancies of 44 pregnant women with SLE.The relationship between lupus flares and pregnant outcomes,and the risk factors for adverse maternal and fetal prognosis were analyzed.T-test,X2 test or Fisher's exact test and Logistic regression were used for statistical analysis.Results① Lupus flares occurred in19(41％)pregnancies(group A) and stable lupus disease was observed in 27(59％) pregnancies(group B) during pregnancy.Compared to pregnancies in patients with stable lupus disease at the conception(n=32),pregnancies in patients with unstable lupus disease at the conception(n=8) had higher lupus flare during pregnancy( 100％ vs 16％,P＜0.05).(②) The common manifestations of lupus flares during pregnancy were lupus nephritis (LN) (11 cases),skin rashes (10 cases),arthritis (7 cases),and the common complication was infection ( 11 cases).(③) The incidence of premature labor,fetal growth retardation (FGR) and fetal loss in group A was 42％,47％ and 26％ respectively,which was significantly higher than that of the group B (7％,15％ and 0 respectively)(P＜0.05).There was no difference in the incidence of preeclampsia,fetal distress and neonatal asphyxia between the two groups ( 16％ vs 7％,16％ vs 19％,5％ vs O,respectively,P＞0.05).The incidence of premature labor and FGR in patients with active LN was higher than that of patients without active LN (55％ vs 11％,64％ vs 17％,respectively,P＜0.05).(④)The binary Logistic regression analysis showed that renal impairment,hypocomplementemia,aPL and serum urea nitrogen level were independent risk factors for premature delivery,FGR,fetal loss and fetal distress.Conclusion(①) Lupus flares during pregnancy increase the incidence of premature labor,FGR and fetal loss.Active LN during pregnancy can increase the incidence of premature labor and FGR.② Renal impairment,hypocomplementemia,aPL and serum urea nitrogen level are associated with adverse fetal outcomes in pregnant patients with SLE.

ObjectiveTo quantitatively assess the effect of Constraint Induced(CI) Movement Therapy on stroke and head injured patients.MethodsNine stroke and head injured patients were applied by CI therapy,which consist of restraint of the unaffected upper extremity in a sling for 14 days combined with 6 hours of training per day of the affected upper extremity. The Carroll test and a Japan upper extremity function test were applied in baseline, pretreatment and posttreatment.ResultsThere were improvement in scores of the Carroll test (P=0.02) and a Japan upper extremity function test (P=0.42)in posttreatment compared with those in baseline.ConclusionsCI therapy is an effective treatment for upper extremity function of stroke and head injured patients.