Objective To provide a method for identifying the seeds of Astragalus membranaceus var.mongholicus(AMM) and A.membranaceus(AM) and to provide basic research data for establishing relevant Standard Operating Practice(SOP) in accordance with GAP of traditional Chinese medicinal materials.Methods The seed morphologic characteristics and microcosmic structures were observed by eyes,light microscopes,and electron microscopes;the seed germination rates of AMM and AM were also compared.Results There was no obvious discrimination of the seed morphology between AMM and AM.There was obvious discrimination on the characteristic of germination-hole,the microcosmic structures of seed-umbilici and seed-coats of seeds between AMM and AM.The hard seed percentage for AMM was higher than that of AM,and its sprouting was not even and sprouting peak appeared later than that of AM.(Conclusion) The seeds of AMM and AM can be identified accuratly with electron microscopes.The patterns of germination-hole,microcosmic structures of seed-umbilici and seed-coats can be used as indices to identify the seeds of AMM and AM.The hard seed percentage and germination characteristics can be used to(identify) the seeds of AMM and AM subsidiarily.

Objective To investigate interventions of vitamin E( VE) on indomethacin-induced gastric mucosal injury. Methods Fifty mice were randomly divided into 5 groups. Normal control group was administrated with saline, others were intragastrically administrated with indomethacin ( 20 mg · kg-1 ) . After 4 hours, mice were intragastrically treated with saline, cimetidine(200 mg·kg-1),VE(25,50 mg·kg-1) once a day for 5 consecutive days. By the end of the treatment,gastric mucosa injury index,serum superoxide dismutase(SOD) activity,and malondialdehyde(MDA) content were determined. ResultsGastric mucosal injury indexs of mice treated with cimetidine,VE at low and high doses were significantly lower than that of the model control and higher than that of the normal control(P<0. 01). VE at high doses showed significantly less mucosal injury than that of the cimetidine group(P<0. 05). Healing-rate of gastric mucosal injury in cimetidine group,VE low-dose and high-dose groups were significantly higher than that of model control group(P<0. 01). SOD activity of VE at low and high doses was significantly lower than that of the model control and cimetidine groups(P<0. 05). The results also revealed that cimetidine and two doses of VE significantly elevated SOD activity and lowered the level of MDA(P<0. 05, 0. 01). Conclusion VE can obviously promote the healing of gastric mucosal injury,especially at high dose,which is better than cimetidine,while,the low dose is comparable with cimetidine.

Objective To compare the clinical efficacies between warm needling and Ibuprofen sustained release capsules (a nonsteroidal anti-inflammatory drug, NSAID) in treating patients with dysmenorrhea in adenomyosis. MethodSixty-five patients with dysmenorrhea induced by adenomyosis were randomized into a treatment group of 33 cases and a control group of 32 cases. The control group was intervened by oral administration of Ibuprofen sustained release capsules, while the treatment group was intervened by warm needling.The intervention lasted 3 menstrual cycles and a 3-month follow-up was studied. The Visual Analogue Scale (VAS), dysmenorrhea symptoms scores and clinical efficacy were compared between the two groups.ResultThe VAS scores after the intervention and inthe first and second months of the follow-up study were significantly different from the pre-treatment score in the two groups (P<0.01); the VAS score of the 3-month follow-up was significantly different from the score before the intervention in the treatment group (P<0.01). There were significant differences in comparing the VAS score after the intervention and in the follow-up study between the two groups (P<0.01), and the treatment group was superior to the control group. The dysmenorrhea symptoms scoresdeclined significantly after the intervention and in the first and second months of the follow-up study in both groups (P<0.01); the dysmenorrhea symptoms score of the 3-month follow-up study decreased in the treatment group and was significantly different from the pre-treatment score (P<0.01). There were significant differences in comparing the dysmenorrhea symptoms scores in the second and third months of the follow-up study between the two groups (P<0.01). The total effective rate was 93.9% in the treatment group, significantly better than 62.5% in the control group (P<0.01).ConclusionWarm needling is effective in easing pain and improving the symptoms of dysmenorrhea in adenomyosis, and can produce a consistent efficacy after the termination of thetreatment; it's superior to NSAIDs in comparing both short-term and long-term treatment efficacies.

BACKGROUND: Previously, there are some researches on studying the effects of hypoxia or hypothermia singly. Nowadays, study on the synthetic action of hypoxia and hypothermia has been attached widely importance.OBJECTIVE: To explore the combination effects of acute hypothermia and hypoxia on superoxide dismutase(SOD) and malondialdehyde in normal rabbits and rabbits with myocardial ischemia.DESIGN: Randomized controlled trial.SETTING, MATERIALS and INTERVENTIONS: Experimental site:Physiological Laboratory of Youjiang Ethnic Medical College. A total of 32 healthy rabbits weighed between 1.8to 2.5 kg were chosen. They were randomly divided into 4 groups each with 8 rabbits: normal group, hypothermia and hypoxia group, ischemic group with normal temperature and oxygen,ischemic group with hypothermia and hypoxia. Normal rabbits and ischemic rabbits were put in fridge at low temperature( -10 ± 2) ℃ and low oxygen (oxygen content of 8.5% ) for one hour and tested activity of SOD and concentration of malondialdehyde in serum.MAIN OUTCOME MEASURES: Comparison of serum SOD and malondialdehyde between normal rabbits and myocardial ischemic rabbits under the acute hypothermia and hypoxia conditions.RESULTS: The SOD level of rabbits in hypothermia and hypoxia group [(371.04 ± 29.96) kNU/L] was much lower than that of normal temperature and oxygen group[ (424.09 ± 22.59) kNU/L] ( t = 5.21, P < 0.01) while the content of malondialdehyde[ (5.58 ± 0. 44) μ mol/L] was greater than that of normal group[ (4.44 ± 1.11) μ mol/L] ( t = 3.21, P < 0.05) . In myocardial ischemia group with hypothermia and hypoxia, SOD was greatly lower than that of myocardial ischemia group with normal temperature and oxygen(t=4.37, P < 0.01) while the malondialdehyde was greatly higher than that of ischemic group with normal temperature and oxygen group(t=2.94, P <0.05).CONCLUSION: The combined effects of acute hypothermia and hypoxia will aggravate the damage to body.

Objective:To design and construct the replication-deficient recombinant adenovirus Ad-siCTGF which can silence the expression of connective tissue growth factor (CTGF) by RNA interference and verified its function. Methods:A specific sequence, which was verified to be able to silence CTGF gene with high efficiency, was cloned into pSES-HUS vector to produce the shuttle plasmid pSES-siCTGF. The plasmid after Pme Ⅰ linearization was cotransduced with pAdEasy into BJ5183 E.coli strains to construct recombinant vector Ad-siCTGF. After linearization treatment with Pac Ⅰ enzyme digestion Ad-siCTGF was transfected into HEK293 cells via liposome mediation. The recombinant adenovirus was packaged. The titer of the Ad-siCTGF was increased after three times of cross-infection. 4T1 cells were infected with the adenovirus. The silencing efficiency was tested by real-time fluorescence quantitative (RFQ)-PCR and Western blotting.Results:Pac Ⅰ enzyme digestion electrophoresis indentified that recombinant adenovirus was successfully constructed. The titer of the recombinant adenovirus Ad-siCTGF was 2.6×10~(10) pfu/mL after amplification and purification. The CTGF mRNA and protein expression in 4T1 cells were decreased by 36.27% and 31.56%, respectively, compared with the control groups.Conclusion:The recombinant adenovirus which can silence the expression of CTGF was successfully constructed. It laid a good foundation for further investigation of the action mechanism of CTGF in tumor cells.

Objective To investigate the blood pressure control effect of captopril sustained-releasetablets based combination, antihypertensive therapy on hospitalized high-risk patients with hypertension in high altitude region. Methods According to the blood pressure,risk factors and combined target organ damage,331 hospitalized patients with essential hypertension were divided into 2 groups and accepted different treatment:low-risk group had monotherapy (n=102) , and night-risk group had captopril sustained-release-tablets based combination antihypertensive therapy (n =229). The discharge blood pressure,extent of SBP/DBP decrease and the compliance rate of discharge blood pressure of two groups were compared. Results The extent of SBP/DBP decrease in hight-risk group was significantly greater than low-risk group (SBP [36. 83 ± 22. 23] mm Hg vs.[28. 74 ±18.71] mm Hg,t=-3. 207,P <0. 05;DBP[22. 04±13. 57]mm Hg vs. [17. 98 ± 13.63] mm Hg,t =-2. 509, P < 0.05). The average discharge blood pressure in both groups reach the standard criterion, but no significant difference was observed between the two groups (SBP [125. 62 ± 14. 74] mm Hg vs. [122. 28 ±13.13]mmHg,t=-1. 962,P>0. 05;DBP[80. 67 ±9. 82]mm Hg vs. [78. 40 ±9. 97]mm Hg,t =-1.910,P > 0. 05). Furthermore we found no significant difference in the control rate of blood pressure between high-risk and low-risk group (72. 06% vs. 71. 57% , x2 = 0.928, P > 0. 05). Conclusion The captopril sustainedrelease-tablets based combination antihypertensive therapy is a reliable treatment in high-risk patients with hypertension from high altitude region,which shows satisfying blood pressure control rate.

Objective To establish a key technological system for spider fibroin gene code tandem connection , vector construction , prokaryotic expression and purification using genetic engineering in order to achieve MaSp 1 heterologous ex-pression in Escherichia coli and its separation and purification .Methods Isocaudarner ligation method was used to connect synthetic spider fibroin gene monomer code in tandem , and a recombinant clone concatemer was obtained .The identified recombinant clones were connected with prokaryotic expression vector pET 28a(+), and then transformed into E.coli BL21 (DE3).After being induced by IPTG for 6 hours, the expression product was identified by SDS-PAGE and Western blot-ting.Engineering bacteria were fermented in high density , and the obtained protein was purified through ammonium sulfate fractionation.Results and Conclusion The expression plasmids of MaSp1concatemers were successfully constructed , and the induced expression genetic engineering MaSp 1 protein was of the expected relative molecular mass .In addition, the pu-rity of the purified protein was above 80%.This study has developed crucial technologies for mass production of genetic en-gineering spider silk proteins .

Objective To harvest pancreatic tissues from rats , prepare decellularized bio-derived pancreatic scaffolds ( DBPS) , and to examine the integrity and biocompatibility of the scaffolds .Methods Normal pancreases were harvested from healthy adult SD rats .DBPS was prepared by perfusing SDS and Triton X-100 through bile duct and the portal vein, respectively.After decellularization, normal pancreatic tissue and DBPS were compared via HE staining , and transmission electron microscopy ( TEM ) . Abdominal wall and subcutaneous implantations were used to compare biocompatibility , and the remain quantity of residual protein and growth factors were determined via enzyme linked immunosorbent assay(ELISA).MTT assay was used to test the scaffolds’ cytotoxicity.The scaffolds were co-cultured with endotheliocyte .Results HE staining and TEM study indicated no residual cells in the DBPS as well as preservation of the complete extracellular matrix .The remain quantity of residual protein and growth factors in ECM was high .The abdominal wall and subcutaneous implantation revealed that DBPS triggered a lower immune response as compared to the control group.MTT assay showed little cytotoxicity .Endotheliocyte assembled and growed with the scaffolds together .Conclusion DBPS are completely decellularized , and exhibit a higher level of biocompatibility in vivo.Using the way of vessels can make the integrity of extracellular matrix to be fully preserves and contain more growth factors .So using vessels way is better than bile duct .

Background Local medical treatment of refractory immunologic keratitis is unsuccessful,and systemic steroids and immunosuppressive agents could cause severe side-effects.Tacrolimus is a potent immunosuppressive drug,it has been proved that topical application of tacrolimus could reduce immunologic inflammation.The safety and efficacy of 0.05％ tacrolimus eye drops for refractory immunologic keratitis has not been described.Objective This study was to evaluate efficacy and safety of 0.05％ tacrolimus eye drops for refractory ulcerative keratitis.Methods A retrospective study was performed.Twenty-one eyes of 17 patients with refractory immunologic keratitis,which had uncontrolled inflammation despite initially treatment including topical steroids and 1％ cyclosporine A,were enrolled,including 11 males and 6 females,with the mean ages of 52 years.Infectious ulcer was excluded by laboratory tests.No systemic disease was found in 11 patients,and Wegener's granulomatosis,rheumatoid arthritis and ulcerative colitis were seen in 1 patient,4 patients and 1 patient respectively before presentation and they were all in remission under conventional systemic therapy.Four patients got binocularly involved and thirteen patients were monocularly involved.Of the 21 eyes,2 eyes with ulcer were ≥ 3 quarters of the limbus,and 19 eyes with ulcer were ≤ 2 quarters.All patients were treated with 0.05％ tacrolimus eyedrops after discontinuing cyclosporine A.The dosage was adjusted according to the severity of inflammation and was gradually tapered when improvement occurred.The corneal lesions were examined under the slit lamp microscope and Heidelberg HRT3 Rostock Cornea Module regularly,and inflammatory cell infiltrations were analyzed with Cell Count(R) software (Heidelberg Engineering GmbH).The safety variables were monitored regularly,including adverse response of eye,tacrolimus blood concentrations measured by chemiluminescent microparticle immunoassay (CMIA) and laboratory examinations of blood routine,blood glucose level,liver and kidney function.Results The patients were treated and followed-up for a mean duration of 18.1 months (range,8-24 months).Corneal ulcer area was obviously reduced 1 month after treatment in 19 eyes,and 2 eyes of 2 cases received anterior lamellar keratoplasty due to progressive corneal destruction despite of tacrolimus therapy.Corneal ulcer was cured 3 months after treatment,and stromal edema and infiltration disappeared 6 months after treatment under the slit lamp microscope.The inflammatory cell densities at lesion zone were (958±329),(858±339),(459±261),(192±124),(98±52),(44±24) and (3±2)/mm2 before treatment and 1 week,1 month as well as 3,6,12,24 months following treatment,respectively,showing a gradually decline as time lapse (F =125.439,P =0.000),and the inflammatory cells were significantly decreased in 1,3,6,12 and 24 months following the administration of 0.05％ tacrolimus eye drops in comparison with that before treatment (all at P =0.000).The therapy duration was 12 months in 9 eyes and 24 months in 12 eyes.Transient irritation sensation occurred in 4 eyes during the treating period.Blood concentrations of tacrolimus were below 1.0 ng/ml in all of the patients.No abnormality was found in laboratory tests.Conclusions The use of 0.05％ tacrolimus eye drops is a safe and effective approach to refractory immunologic keratitis.

Objective To investigate the differentiation of neuroendocrine component (NEC) in colorectal adenocarcinoma in relation to its significance by comparing the outcome between patients with or without NEC.Methods The paraffin sections from patients with pathologically confirmed colorectal adenocarcinoma were retrospectively collected and screened for those with NEC by morphological examination and immunohistochemistry with neuroendocrine markers.Control patients (n=54) without NEC were selected from colorectal cancer database and 2: 1 matched on the basis of clinical features with NEC positive patients (n=27).Relative analysis was performed between two groups.Results With a median follow-up of 72 months,the 5-year disease free survival was 58.0% (16/27) in NEC positive group and 79.1% (43/54) in control group (P=0.036).Similarly,the 5-year cancer-specific overall survival was significantly lower in NEC positive group than in control group (58.3% versus 81.1%,P=0.037).Cox regression showed that the 5-year cumulative risks of disease recurrence and cancer-caused death in NEC positive patients were 2.38 and 2.41 times higher than those in control patients,respectively.Conclusions NEC appears to bear a poor prognosis in patients with colorectal adenocarcinoma.