Objective To clone amastin coding genes from different strains of Leishmania parasites. Methods Using amastin cDNA sequence as the reference, dbEST data base established by National Center Biotechnology Information (NCBI), USA, was searched by BLAST tool. A 309 bp DNA fragment of Leishmania major was found and used as the probe for the screening of a DNA library. The amastin gene of Leishmania major Abdou was cloned and sequenced. Specific primers were designed and amastin genes for Leishmania mexicana WR972, Leishmania brizeliensis and Leishmania amazonensis joseph were amplified by polymerase chain reaction. Results The amastin genes from 4 strains of Leishmania parasites were cloned and sequenced. It was found that all 4 amastin genes contained unique open reading frame of 552 bp and encoded amastin protein of 183 amino acid residues. Conclusion The amastin genes of 4 strains of Leishmania parasites were successfully cloned.

Abstract?AIM: To analyze the effects of femtosecond laser assisted cataract surgery ( FLACS ) in the treatment of cataract and its effect on prognosis.?METHODS:Forty-two cases (42 eyes) of patients with cataract who were treated in the Department of Ophthalmology in our hospital between January 2012 and December 2014 were selected as the study objects. According to the order of treatment, they were divided into control group and observation group, 21 cases in each. The control group was treated with traditional phacoemulsification cataract surgery ( PCS ) . On the basis, the observation group was treated with femtosecond laser. The effective phacoemulsification time (EPT), cumulative dissipated energy (CDE), fluid flow and monitored pressure of the two groups were recorded.The rate of corneal endothelial loss and the situation of Tyndall phenomenon were statistically analyzed.The two groups were followed up for 1a.The long-term visual acuity recovery was observed.The best corrected visual acuity ( BCVA ) was recorded, and the long-term complications were statistically analyzed.?RESULTS: 1 ) The total response rate in observation group was 95% while in control group was 90% ( P>0.05);2) the surgery time of the observation group was longer than that of the control group ( P<0.05) but EPT was shorter than that of the control group.CDM and liquid flow were less than those of the control group ( P<0.05 ); 3 ) at 1d after surgery, there was no significant difference in intraocular pressure between the two groups (P>0.05); the rates of Tyndall phenomenon and corneal endothelial loss in the observation group were lower than those in the control group ( P<0.05);4) BCVA of the two groups at different time after surgery were significantly higher than that before surgery (P<0.05). However, at 1d, 3mo, 6mo and 1a after surgery, BCVA of the observation group was better than that of the control group ( P <0.05 ); 5 ) the incidence of complications in the observation group after surgery (14%) was lower than that in the control group (43%) (P<0.05).?CONCLUSION: The surgical effects of FLACS in the treatment of cataract are good.After surgery, the visual acuity of patients is improved significantly and the incidence of postoperative complications is low. However, the surgery time is long and cost is high, so it is difficult to popularize.

Objective To evaluate clinical effects of integrated traditional Chinese and western medicine therapy for epilepsy after stroke. Methods Literatures about the treatment of epilepsy after stroke with the combination of traditional Chinese medicine and western medicine which come from the database such as CNKI, VIP, WanFang and Pubmed, were studied and all the randomized controlled trials on comparing therapeutic effects of treating epilepsy between combined treatment of TCM and western medicine and western medicine were enrolled. The Cochrane Collaboration's software RevMan 5 was used for meta-analysis. Results 7 articles were included in the study, there is homogeneity among these articles(P>0.05), The fixed effect model was used for the meta analysis. the combined results were OR=5.17, 95%CI(3.02, 8.82), the overall effect of test Z=6.01, P<0.01, the difference between the treatment group and the control groups was significant, that means the overall effect of the combination of traditional Chinese medicine and Western medicine treatment was better than western medicine treatment; Funnel plot graphic displayed asymmetrically, which indicated the literatures publication bias. Conclusion The current data indicate that the curative effect of TCM and west medicine combined treatment is better than pure western medicine therapy for epilepsy after stroke, but as the literature publication bias, clinical studies need more high quality large samples to prove this conclusion.

BACKGROUND: The decrease of activity level of choline acetyltransferase (ChAT) and reduction of membrane protein of synaptic vesicles are the main biochemical indexes in Alzheimer disease. Traditional Chinese bushen yizhi formula is of the effects of strengthening the kidney, replenishing qi, generating marrow and nourishing the brain, its medicine-containing serum can effectively protect these changes resulting from amyloid protein.OBJECTIVE: To investigate whether the medicine-containing serum of bushen yizhi formula can improve the pathological injury of neurons induced by amyloid protein metabolites in Alzheimer disease, thus to estimate the therapeutic effect of bushen yizhi formula on Alzheimer disease.DESIGN:A randomized and controlled trial. SETTING:Institute of Neuroscience, Second Affiliated Hospital, Guangxi College of Traditional Chinese Medicine, Nanning 530011, Guangxi Zhuang Nationality Autonomous Region, China; Encephalopathic Institute of Guangzhou University of Traditional Chinese Medicine.MATERIALS:The experiment was conducted from September 2003 to March 2004 at the Research and Development Center of New Drugs of Guangxi College of TCM. Totally 60 healthy Wistar rats of clean grade,three months, were at random divided as blank control and bushen yizhi groups, with 30 in each group. The bushen yizhi formula consisted of Fructus Lycii, Fructus Cnidii, Radix Gingeng, Radix Polygoni Multiflori,Cortex Moutan Radicis, Borneolum, etc. The medicine was prepared before use as 0.176 g/Ml liquid(including 1.13 g/Ml rude drugs).METHODS:① Each of the rats in bushen yizhi group was by garage given 1.0-1.5 Ml prepared liquid per day for consecutive 30 days. Within 4hours after the last administration the serum was isolated at 5 000 r/min for 5 minutes, filtered aseptically, the compliment was inactivated at 56 ℃,then the medicine-containing serum was obtained. The rats in blank control group were given saline of the same volume, and the serum was prepared with the same method. ② For NG108-15 cells, the concentration of amyloid protein 25-35 segments in the culture liquid was 5 μ mol/L, or 1-42 segments, 200 nmol/L, was suitable. ③ NG108-15 cells was first in cubated for a day in CO2 incubator with Dulbecco-improved Eagle medium of 0.1 molarity at 37 ℃, then 5 μmol/L amyloid protein 25-35 segment or 200 nmol/L segment 1-42 was added to continuously incubate for a day, after that the cell models of Alzheimer disease was ready. Then 50 g/L medicine-containing serum or Dulbecco-improved Eagle medium of blank control group serum was taken to replace the original culture liquid, and the amyloid protein segments of the above-mentioned concentration was still added for maintaining the cell models of Alzheimer disease, after 5 days'differentiation incubation, the indexes were detected with those in non-amyloid protein as control. ④ Western blotting, radioimmunoassay and electrophysiological examination were used to investigate the ChAT activity,synapsin protein level and rate of functional synapse formation of NG108-15cells in Alzheimer disease after treatment with the medicine-containing serum of bushen yizhi formula.MAIN OUTCOME MEASURES :The effects of the medicine-containing serum of bushen yizhi formula on ChAT activity, synapsin protein level,rate of functional synapse formation of NG108-15 cells and frequency of miniature plate potential in NG108-15 cells.RESULTS:Totally 60 rats involved all entered the final result analysis.① The effect of the medicine-containing serum of bushen yizhi formula on ChAT activity: In the conditions when there was no amyloid protein or the final concentration of amyloid protein 1- 42 was 200 nmol/L, the ChAT activity in bushen yizhi group was obviously higher than that in blank control group [(1651.2±134.5), (1336.1±268.2), ( 586.1±223.4),(1290.7±381.5)μmol/(g·h); P ＜ 0.05] . ② The effect of the medicinecontaining serum of bushen yizhi formula on synapsin protein level: The absorbance values of synapsin Ⅰa and Ⅰb, in the conditions when there was no amyloid protein, amyloid protein 25-35 and 1-42, were respectively 5.02, 1.36 and 2.46 in bushen yizhi group; and 3.18, 0.57 and 0.71 in blank control group. The values in bushen yizhi group were respectively 1.6,2.4 and 3.5times those in blank control group. ③ The effect of the medicine-containing serum of bushen yizhi formula on functional synapse formation of NG108-15 cells: In the conditions when there was no amyloid protein or amyloid protein 1-42, the levels of functional synapse formation in bushen yizhi group wereall higher than those in blank control group [(90.6±6.0)%, (63.2±17.0)%, (58.0±13.1)%, (34.2±13.0)% ;P＜0.05].④ The effect of the medicine-containing serum of bushen yizhi formula on frequency of miniature plate potential in NG108-15 cells: In the conditions when there was no amyloid protein or amyloid protein 1-42, the frequencies of miniature plate potential in bushen yizhi group were all higher than those in blank control group [(9.28±4.1), (5.48 ±5.14), (5.55±5.85),(3.05±4.46)/min; P ＜ 0.01, P＜ 0.05].CONCLUSION:In the conditions when there existed no amyloid protein or amyloid protein, the medicine-containing serum of bushen yizhi formula can raise ChAT activity, not only correct the decrease of cellular synapsin caused by amyloid protein, but also raise the expression of synapsin protein, the rate of functional synapse formation and neurotransmitter release under the condition of no amyloid protein. It was shown that the prescription can antagonize the pathological development of Alzheimer disease, and play the therapeutic effect through enhancing the release power of neurotransmitter.

Objective To investigate the cell cycle and mechanisms of HCC Bel-7404 cell line by treatment of extract of centipede ( ECP). Method Bel-7404 cell line was cultured in vitro. ECP was applied to the interference of the growth of Bel-7404 with different drug concentration. Cell cycles and apoptosis ratios of ECP group were detected by flow cytometry (FCM). To investigate the expression of apop-tosis related genes XIAP and Bax, we also detect HCC Bel-7404 cell line after 48 hours treated with ECP by immunohistochemical method. Result The results of FCM displayed that ECP in treatment group mainly retard cell cycle phase in G0/G1 after 48 hours. The ratios of G0/ G1 .proliferation index (PI) and apoptosis ratios were significantly different between treatment group and control group when the concentrations were different. It was also significantly different between treatment group and control group when the same concentration of ECP was used. It showed that XIAP gene expression decreased gradually while Bax gene expression increased with the increase of ECP concentration, and these were statistical significance in contrast to control group ( P <0.05). Conclusion ECP mainly retarded cell cycle phase of Bel-7404 in G0/G1, suppressed the proliferation, and could induce it to apoptosis. The inhibitory effect was time - concentration dependent. The mechanisms were due to promote Bax gene and inhibit XIAP gene expression in HCC Bel-7404 cell line.

Objective:To investigate the immune mechanism of 5-fluorouracil(5-Fu) and Octerotide(Oct)in the treatment of severe acute pancreatitis(SAP)and it’s kidney injury.Methods:55 male SAP rats induced by retrograde infusion of 5% sodium taurocholate into bilopancreatic duct were randomly divided into 3 groups:Normal saline group(n=17)were treatmented with normal saline infusion,Oct group(n=18):The first dose of Oct was 2?g/100g iv,then 0.2?g/(100g?h)continuous injection,5-Fu group(n=20):The dose of 5-Fu was 2mg/h?12h.Rats from each group were killed at 12h after opertation.The plasma levels of tumor necrosis factor-?(TNF-?),interleukin-1(IL-1)were assessed by ELISA method.Thromboxan B_2(TXB_2)and prostaglandinE_2(PGE_2)were assessed by radioimmunoassay method.The expression of TNF-? mRNA was detected by RT-PCR.The apoptosis rate of rat kidney tissue were detected by TUNEL method.Apoptosis rate of NRK cell treated by serum of different groups were assessed by flow cytometry methods.Results:The plasma level of TNF-?、IL-1 and TXB_2 in 5-Fu and Oct group were much lower than that in normal saline group.The expression of TNF-? mRNA of kidney tissue decreased significantly after the SAP was treated by 5-Fu or octreotide.The apoptosis rate of NRK cell disposed by normal saline group,oct group,5-Fu group and control group rat plasma were (38.67?11.4)%,(20.4?18.4)%,(10.5?11.0)% and (1.7?2.2)% respectively(P

Objective To assess the role and its tecnhique superiority of 64-slice spiral CT angiography(CTA) in lower limb arterial diseases.Methods MSCT studies in 33 cases suspected diseases of lower limb arteries was performed,the helical scans covered the extent of renal artery to the artery of the lower extremity.3D reconstruction of vessel was done in Wizard workstation.The manifestations of MSCTA were compared with that of conventional angiography . Results Of the 693 segments of arteria , 683 were assessable on both DSA and CTA.Using the DSA images as the standard reference,the sensitivity,specificity and accuracy of CTA were 97.6%(95% CI,94.9%~100.7%),98.9%(95% CI,98.0%~99.8%),98.7%(95% CI,97.9%~99.5%)in detecting the segments occluded . With the regard the detection of segments that had more than moderate stenosis , the sensitivity , specificity and accuracy of MIP were 98.9% ( 95% CI , 97.4%~100.4%),99.4%( 95% CI, 98.7%~100.1%) and 99.3%(95% CI, 92.4%~99.9%).Conclusion In assessment of diseases of lower limb arteries with 64-slice spiral CT angiography is equal to DSA basically, it is a highly accurate and non-invasive angiography in diagnosing arterial diseases of the lower extremities.

AIM: To evaluate the genotype, muscle histopathology and ultrastructure in dko mice. METHODS: Dystrophin/Utrophin-deficient double knockouts (dko) mice were obtained from university of Oxford, UK. Genotype of filial generation of heterozygote was evaluated by PCR-SSP. HE staining and fluorescent immunohistochemistry by SABC-Cy3 were used to detect striated muscle of dko mouse, and the muscle ultrastructure was observed by transmission electron microscope(TEM). RESULTS: In 112 filial generation mice, there were 28 mdx (25.0%), 26 dko (23.2%) and 58 heterozygote (51.8%), which coincided with the law of Mendelian inheritance. HE staining showed that the myocytes were not very uniform, there were phenomenon of round outline, centrally nucleated fibers, widening interspace, inflammatory cell infiltration and connective tissue proliferation in dko mice. There were no any immunofluorescent expression of dystrophin and utrophin in sarcolemma in dko mice. TEM showed sarcolemma breakage, separation and edema, and loose myofibril texture, inflammatory cell infiltration and connective tissue proliferation in dko mice. CONCLUSION: PCR-SSP is a very quick and accurate way for genotype evaluation of filial generation. The pathophysiology of dko mouse was very similar to Duchenne muscular dystrophy (DMD), and dko mouse is an ideal animal model for study of DMD clinical therapy.

AIM: To observe amelioration of motor function in a Duchenne muscular dystrophy (DMD) mouse model ( dko mice) after transplantation of bone marrow mesenchymal stem cells (MSCs). METHODS: Passage fifth MSCs cultured in vitro were transplanted into dko mice by tail vein, motor functions of experimental mice and matched control mice, including traction, rotating rods, rotated wheel, upside down, turning over and walking (all were recorded by Sony digital camera) were tested 15 weeks after transplantation. The fluorescent expression of dystrophin and utrophin in gastrocnemius muscle tissue of dko mice was detected by SABC-Cy3, and average optical density of positive fibers was calculated. RESULTS: MSCs grew in colony over passage third, and there was low immunologic reaction by vein transplantation. There was dystrophin and utrophin fluorescent expression in sarcolemma of dko mice 15 weeks after transplantation, but no any fluorescent expression in controls. There was significant difference in fluorescent average optical density of positive fibers between two groups ( P