Objective:To clone and express survivin gene in mammalian cells.Methods:Coding sequence of survivin gene was amplified from HepG 2 cell mRNA by RT-PCR and then cloned into eukaryotic vector pEGFP-C1.The vector with survivin gene was transfected into HepG 2 and SMMC-7721.The expression of survivin was detected by GFP and Western blot.Results:The recombinant plasmid pEGFP-C1-survivin was successfully constructed and expressed in HepG 2 and SMMC-7721.Conclusion:The result shows expression of the anti-apoptosis gene survivin in mammalian cells.

Objective:To construct the plasmid containing short hairpin RNA(shRNA) of green fluorescent protein(GFP) to suppress the expression of GFP.Methods:A 20bp reverse repeated motif of GFP target sequence with 4bp spacer were synthesized,and inserted into pTZU6+1 to generate the pshRNA—GFP plasmid;pshRNA—GFP and pEGFP-C1 or pTZU6+1 and pEGFP-C1 were cotransfected into mammalian cells to detect effect of GFP expression separately.Results:pshRNA—GFP suppressed the GFP expression by 70%～85% in mammalian cells.Conclusion:The result shows that the short hairpin RNA of GFP can efficiently suppress its expression in HepG2 and SMMC-7721.

Objective To investigate the application value of cataract extraction surgery in the treatment of primary angle-closure glaucoma and cataract.Methods 132 patients of primary angle-closure glaucoma and cata-ract were randomly selected,these patients were divided into lensectomy treatment group(A group,n=66) and laser peripheral iridotomy treatment group(B group,n=66) according to the treatment method.The visual acuity,intraocu-lar pressure,anterior chamber depth,and complications of the two groups were analyzed statistically.Results In A group,the visual acuity <0.1 was in 0 case,0.1-0.3 in 12 cases,>0.3-0.4 in 20 cases,>0.4 in 34 cases, accounting for 0.00%,18.18%,30.30%,51.52%.In B group,the visual acuity <0.1 was in 5 cases,0.1-0.3 in 14 cases,>0.3-0.4 in 19 cases,>0.4 in 28 cases,accounting for 7.58%,21.21%,28.79%,42.42%.A group of visual acuity <0.1 proportion of patients was significantly lower than B group(χ2 =5.02,P<0.05) ,the propor-tion of patients >0.4 was significantly higher than B group(χ2 =7.38,P<0.05),but the differences of the propor-tion for 0.1-0.3,>0.3-0.4 vision between the two groups were not significant(χ2 =2.71,4.61,all P>0.05). The differences of the intraocular pressure and anterior chamber depth before surgery between the two groups were not significant(t=1.532,1.447,all P>0.05),the anterior chamber depth after surgery in group A was significantly lon-ger than B group(t=2.571,P<0.05),but the difference of IOP between the two groups was not significant(t=1.032,P>0.05).A group had corneal edema in 1 case,a transient high IOP in 4 cases,2 cases in hyphema,the postoperative complication rate was 10.61%.B group had corneal edema in 1 case,a transient high IOP in 4 cases, 2 cases in hyphema,the postoperative complication rate was 10.61%,the incidence rate of postoperative complica-tions in A group was significantly lower than B group(χ2 =9.35,P<0.05).Conclusion The application value of cataract extraction surgery in the treatment of primary angle -closure glaucoma and cataract is higher than laser peripheral iridotomy,which can more effectively improve visual acuity, prolong anterior chamber depth, reduce inci-dence of complications of patients,so it is more safe and effective,and worthy of promotion.

OBJECTIVE:To prepare different compound scald preparations and to investigate their repairing effects for deep Ⅱ? scald in rabbits.METHODS:The deep Ⅱ? scald model was established in depilated rabbits by infrared radiation.Then the rabbits in the trial groups were treated with the same drug at different dosage and different forms of medication:compound scalded gels,compound scalded ointments and the compound scalded films,with Moist Exposed Burn Ointment as positive control and 0.9% NaCl aqueous solutions blank.After a certain time of treatment,the contraction of the scald skin was observed and the average value of the diameter of the scald skin was calculated.RESULTS:In three trial groups and Moist Exposed Burn Ointment positive control group,no scald infection occurred,and the healing surfaces of the scalds were all smooth and even.However,the healing time was different between 3 trial groups and Moist Exposed Burn Ointment positive control group,with compound scalded gels showing the best efficacy in that the scald surfaces were free from infection,the healing surfaces were smooth and even,and the contracted diameter of the scalded skin was significantly lower than in other groups,showing significant differences as compared with other groups(P

The mechanism of invasion and metastasis of colorectal carcinoma was associated with many aspects, including correlative genes such as k-ras,c-met,nm23,c-SRC,HES-6 and P107; immunoreaction; angiogenesis; correlative molecules interacting with extracellular matrix,nitric oxide. The paper summarized the current development in this field.

Objective: To study the role of membrane-cytoskeleton linker Ezrin in the growth and metastasis of human colorectal carcinoma (CRC) cell lines. Methods: Human CRC cell lines Lovo and SW 480 were cultured. Three pairs of small interfering RNA (siRNA) targeting Ezrin were transfected into the CRC cells. Western blotting was used to detect the transfection rates so as to screen the most effective siRNA which had been transfected into the CRC cells. Real-time PCR and Western blotting were used to detect the downregulation of Ezrin mRNA and protein at different time points. We observed the biological behaviors of CRC cells when the expression level of Ezrin was the lowest. CCK-8 method was used to detect the proliferation of the cells. Flow cytometry was used to detect the cell cycle and apoptosis. Transwell test was used to detect the invasion and migration ability of the transfected CRC cells. Results: Real-time PCR and Western blotting results revealed that Ezrin siRNA notably down-regulated Ezrin expression at both mRNA and protein levels. Down-regulation of Ezrin expression distinctly decreased the proliferation rates of the two cell lines. After RNAi treatment, the cell proportion in S phase decreased slightly and the cell proportion in G1 phase increased slightly in the two cell lines (P > 0.05). The apoptotic index increased in the three siRNA transfected groups compared with the control group (P < 0.05). The migration and invasion ability of CRC cells significantly decreased after siRNA interference (P < 0.01). Conclusion: Ezrin plays an important role in the process of proliferation, migration, and invasion of CRC cells. It has the potential to become a new target to inhibit tumorigenesis and development of CRC.

Objective: To evaluate the in vivo pharmacokinetics of strychnos alkaloids [brucine, total alkaloids in Strychni Semen (TASS), and optimal total alkaloids in Strychni Semen (OTASS)] by ig administration to rats. Methods: The rats were ig administered with strychnos alkaloids and their blood samples were taken. Huperzine A was used as an internal standard. The brucine in plasma of rats was detected by HPLC method. The compartment model was fitted and the pharmacokinetic parameters were calculated by DAS1.0 program. Results: The pharmacokinetic analysis showed that brucine behaved as a two-compartment model in the three solutions after ig administration in rats. Compared with brucine monomer administration, the solution of TASS and OTASS could obviously increase the absorption of brucine in vivo, but other pharmacokinetic parameters had no significant difference. Conclusion: After ig administration with strychnos alkaloids, the other alkaloids in Strychni Semen could promote the absorption of brucine.

Humans ; Malocclusion ; therapy ; Molar, Third ; growth & development ; pathology ; Orthodontics, Corrective ; Tooth Eruption ; Tooth Loss ; pathology ; Tooth Movement Techniques

strains that could produce raw starch-digesting glucoamylase were isolated from soil and mildewed rice.The highest raw starch-digesting glucoamylase activity strain named OR-1 was identified as Rhizopus.sp.The raw starch-digesting glucoamylase activity of the strain is 90U/mL.Through UV and NTG mutagenesis,the raw starch-digesting glucoamylase activity raised to 200U/mL and 325U/mL respectively.The RDA were 70% and 65% respectively.

Aim To study the sensitivity of human gastric cancer BGC-823 cells to paclitaxel after trans-fection of SHIP2 ( The SH2 domain containing inositol 5-phosphatase 2 ) cDNA. Methods Apoptotic cells were determined by the propidium iodide method using flow cytometry. The levels of protein and mRNA ex-pression were measured by Western blot analysis and qRT-PCR, respectively. pCMV6-SHIP2 plasmid and empty vector were transiently transfected into BGC-823 cells, respectively. Stable cell lines were established after infecting BGC-823 cell with GV112-Puromycin and GV112-Puromycin-Bim( Bcl-2 interacting mediator of cell death) lentivirus particles. pCMV6-SHIP2 plas-mid was transiently transfected into the stable cell lines. Results BGC-823 cells were relatively insensi-tive to paclitaxel compared with SGC-7901 cells. The apoptotic rate was only (25. 6 ± 1. 6)% after the treat-ment with 0 . 3 μmol · L-1 paclitaxel for 48 h in BGC-823 cells. The expression levels of Bim protein and mRNA in BGC-823 cells treated with paclitaxel at dif-ferent time points were not significantly changed. The expression of Bim protein was increased after transfec-tion of pCMV6-SHIP2 plasmid, and the apoptotic rate was up to ( 50. 8 ± 0 . 9 )% in BGC-823 cells treated with paclitaxel for 48h. The expression of Bim protein was significantly inhibited after infecting with GV112-Puromycin-Bim lentivirus particles. The apoptotic rate of infected BGC-823 cells was only ( 27. 6 ± 1. 6 )%after treatment upon paclitaxel for 48h. Conclusion Overexpression of exogenous SHIP2 can increase the expression of Bim, induce apoptosis and enhance sen-sitivity of BGC-823 cells to paclitaxel.