Objective:To optimize the formula of 5-aminosalicylic acid enema in situ gel. Methods:5-Aminosalicylic acid ene-mas in situ gel was prepared using a cold dissolution method with carbomer as the gel matrix and xanthan gum as the thickener. A 32 full factorial design was used to investigate the effects of the concentrations of carbomer and xanthan gum on the viscosity before and af-ter the gelling, duration of inversion tube and sedimentation rate. Response surface methodology was used to optimize the formula. Re-sults:The quantitative relationships between the two factors and the four evaluation indices were obtained. The optimum formula was as follows:the concentration of carbopol and xanthan gum in the enema was 0. 7% and 0. 15%, respectively. The viscosity before and af-ter the gelling was 500-1 000 mPa·s and 2 200-2 700 mPa·s, respectively. The duration of inversion tube test was 40-80 min and the sedimentation rate was more than 98. 5%. Conclusion:The multi-objective simultaneous optimization of the formula of 5-aminosal-icylic acid enema in situ gel is accomplished by factorial design and response surface methodology.

Objeetive To explore the value of antisense imaging of 99Tcm-labeled ASON targeting mouse double minute 2(MDM2) mRNA for the diagnosis of human prostate cancer.Methods The ASON targeting MDM2 mRNA and the mismatched oligonucleotide (ASONM) were synthesized and radiolabeled with 99Tcm using the bifunctional chelator HYNIC.The labeling efficiency and radiochemical purity were investigated.Animal models of nude mice bearing human prostate cancer LNCaP were established and divided into 3 groups with 10 mice in each group.99Tcm-HYNIC-ASON,99Tcm-HYNIC-ASONM (study groups) and 99TcmO4-(control group) were injected at the dose of 7.4 MBq through the tail vein,respectively.Tumor imaging was acquired with SPECT and the tumor-to-muscle (T/M) ratio was measured.The data was compared by one-way analysis of variance.Results The labeling efficiencies of ASON and ASONM were (65.15± 2.05) ％ and (64.93±2.18) ％,respectively.Their radiochemical purity was greater than 90％.At 1,4 and 10 h post injection,the T/M ratios of 99Tcm-HYNIC-ASON group were 3.217±0.125,3.749± 0.201 and 4.028±0.186,and those of 99Tcm-HYNIC-ASONM group were 1.579t0.128,1.715±1.140 and 1.683±0.139,and control group 2.146±0.132,1.847±0.124,1.528±0.152,respectively.The T/M ratios in control group and 99Tcm-HYNIC-ASONM group were significantly lower than those in 99Tcm-HYNICASON group at 1,4 and 10 h,respectively (F=213.37-235.41,t=3.527-4.738; all P＜0.01).The T/M ratios of 99Tcm-HYNIC-ASONM group and control group were not significantly different at 1,4 and 10 h (t=2.154,2.287 and 2.236,all P＞0.05).Conclusion The antisense probe of MDM2 can accumulate specifically in prostate cancer tissue in animal models,which might be useful as a non-invasive genetic tool for the early diagnosis of prostate cancer.

Objective To investigate the effect of mouse double minute 2 (MDM2) mRNA ASON and mismatched oligonucleotide (ASONM) radiolabeled with 99Tcm on target gene expression in LNCaP cells.Methods The ASON and ASONM targeted to MDM2 mRNA were synthesized and radiolabeled by 99Tcm with the bifunctional chelator of HYNIC.The labeling efficiency,radiochemical purity,stability and molecular hybridization activity were investigated.The different concentrations of 99Tcm-HYNIC-ASON (0,100,500 nmol/L) and 99Tcm-HYNIC-ASONM (500 nmol/L) coated with lipofectamin 2000 were incubated with prostate cancer cells for 24 h,then RT-PCR and Western blot were carried out to assay the MDM2,p53 mRNA and the corresponding protein level.The variables of RT-PCR and Western blot were analyzed using one-way analysis of variance and q test.Results The labeling efficiency of ASON and ASONM were (65.15± 2.05)％ (n=5) and (64.93±2.18)％ (n=5),respectively.The radiochemical purity were both more than 90％.99Tcm-HYNIC-ASON had a good stability and could hybridize to the sense oligonucleotide (SON).The contents of MDM2 mRNA in 0,100,500 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups were 0.458±0.035,0.250±0.026,0.174±0.032,0.463±0.033,respectively,and there were significant differences between each 2 groups except between 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups (F=33.69,q =24.32-91.45,all P＜0.01).The average density of MDM2 protein in the 4 groups were 90.712±3.042,71.218±2.915,32.775±3.062,88.121±2.710,respectively (F=235.93,q=6.43-19.14,all P＜0.01; except 0 nmol/L99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).The contents of p53 mRNA in the 4 groups were 0.185±0.046,0.203±0.040,0.213±0.027,0.163±0.049,respectively(F =2.18,P＞ 0.05).The average density of p53 protein was 33.865 ± 2.213,70.445±2.180,99.025±3.012,38.351±3.271,respectively (F=53.98,q =3.32-6.74,all P＜0.01 ; except 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).Conclusions The MDM2 antisense probe can accumulate in the prostate cancer cells,and specially hybridize to the MDM2 mRNA and inhibit target gene expression.This novel molecular probe has a promising potential for the diagnosis of prostate cancer at gene level.

Objective To explore the protective effects of dipeptidyl peptidase-4 inhibitor (DPP-4I) on AD-like neurodegenerative changes and its mechanism. Methods The human neuroblastoma cell line SH-SY5Y on the logarithmic phase was divided into six groups:control group (CON group, treated with PBS contained 1‰DMSO for 12 h), wortmannin intervention group (W group, treated with 0.03 μmol/L wortmannin for 12 h), DPP-4I intervention group (DPP-4I group, treated with 10μmol/L DPP-4I for 12 h), both DPP-4I and wortmannin intervention group (DPP-4I+W group, pre-treated with 10 μmol/L DPP-4I for 2 h, then 0.03 μmol/L wortmannin for 12 h), DPP-4I, wortmannin and Ex9-39 intervention group (DPP-4I+W+Ex9-39 group, pre-treated with 10μmol/L Ex9-39 for 2 h, then 10μmol/L DPP-4I for 2 h followed by 0.03μmol/L wortmannin for 12 h), and Ex9-39 intervention group (Ex9-39 group, treated with 10μmol/L Ex9-39 for 12 h). MTT assay was used to detect the cell vitality. Western blot assay was used to detect the level of total tau protein (tau-5) and phosphorylated tau at different sites (pSpS199/202, pT231 and pS396), the level of phosphorylated neurofilaments (NF-H, NF-M) and phosphorylation of critical enzyme in PI3K/Akt/GSK-3β signaling pathway. Results (1) The cell vitality decreased, the levels of pSpS199/202, pT231, pS396 and NF-H/M increased significantly in W group than those in CON group. However, comparing with CON group, the above mentioned parameters reversed in DPP-4I group. Comparing with W group, the cell vitality increased and phosphorylated levels of above mentioned indices were decreased in DPP-4I+W group. (2) The cell vitality showed a decline trend while the levels of phosphorylation tau at three different sites and NF-H/M were higher in Ex9-39 group than those in CON group. Comparing with DPP-4I+W group, the results of the phosphorylated levels showed the same changes in DPP-4I+W+Ex9-39 group. (3) Comparing with CON group, the expression levels of phosphorylated PI3K, Akt and GSK3β increased significantly in DPP-4I group, while those decreased in W group. Additionally, the expression levels of phosphorylated PI3K, Akt and GSK3β were significantly increased in DPP-4I+W group than those in W group. Conclusion DPP-4I can enhance the level of GLP-1 and activate PI3K/Akt/GSK-3βinsulin signaling pathway to improve the hyperphosphorylated tau and NFs induced by wortmannin, and to protect AD-like neurodegeneration.

Objective In order to know the effect of heating infusion for premature infants to pro-mote their restoration of physiological function. Methods Divided 160 premature infants who need infu-sion into the experimental group and the control group randomly, there were 80 cases in each group. Normal temperature infusion was used in the control group, while heating infusion was used in the experimental group, compared the body temperature, pluse, respiration, blood pressure, oxygen saturation and the blood ghcose between the two groups. Results There were significant difference about the indexes which had mentioned above between the two groups. Conclusions Heating infusion can effective promote the reha-bilitation of physiological function of premature infants.

Objective To evaluate the detection rate and the efficiency of the modified crushing-cercariae escaping method.Methods The detection rates of the modified crushing-cercariae escaping method and the crushing methods were compared by using a double-blind control experiment with the latter as a gold standard.meanwhile the number of the cercariae was quantified.The efficiency of the two methods aforementioned and the cereariae escaping method were compared in field.Results The detection rate of the modified crushing-cercariae escaping method was 100%.the average number of cercariae in each infected snail was (4 778±1 157);and the number in certain volume of water sample was positively correlated with the number of infected snails.The efficiency of the modified menthed Was 18.2 times and 17.3 times as high as those of the crushing method and cereariae escaping method,respectively.Conclusions The modified crushing-cercariae escaping method Can detect the infected snails quicky and Can quantify the number of infected snails and cercariae,and is suitable for the detection of infected snails in large number.

Objective To explore the mechanisms of integrin β1 on connective tissue growth factor(CTGF)-induced proliferation,migration,change of cytoskeleton of pulmonary arterial smooth muscle cell(PASMC) in vitro,and to investigate the effects of CTGF-integrin β1 signal pathway on pulmonary vascular remodeling in pulmonary arterial hypertension (PAH).Methods Pulmonary artery smooth muscle cells of SD rats were cultured in vitro.WST-1 assay was used to detect the effects of anti-integrin β1 antibody on CTGF-induced proliferation of PASMC.Transwell chambers were used to observe the effects of anti-integrin β1 antibody on CTGF-induced migration of PASMC.The cytoskeletal rearrangement was observed with coomassie brilliant blue R250 staining and Confocal Lasar Scanning Microscopy (CLSM).Results Different concentration of anti-integrin β1 antibody could inhibit the proliferation of PASMC induced by CTGF,which presents concentration dependent pattern (P ＜ 0.05).The higher the concentration of anti-integrin β1 antibody,the more severity the proliferation of PASMC induced by CTGF was inhibited.and inhibition rate of PASMC proliferation was the highest at 72 hours.Anti-integrin β1 antibody(15 mg/L) decreased significantly the number of PASMC passing through Transwell induced by CTGF,compared with CTGF group (P ＜ 0.01).Meanwhile,antiintegrin β1 antibody could change cytoskeletal rearrangement of PASMC induced by CTGF.Conclusions Integrin β1mediates the proliferation,migration,cytoskeletal rearrangement of PASMC induced by CTGF.The CTGF-integrin β1signal pathway may play a key role in proliferation,migration,cytoskeletal rearrangement PASMC.

BACKGROUND: Anterior cruciate ligament (ACL) injury directly impacts the individual's lower limb function. It is very important for althelets whether rehabilitation training after ACL reconstruction is reasonable, effective and can restore the normal function of the knee joint.OBJECTIVE: To explore the effect of rehabilitation training on graft healing and knee joint function recovery after ACL reconstruction. METHODS: An elite rugby player was enrolled and subjected to phased rehabilitation training after ACL reconstruction. MRI examination was done before and after rehabilitation training. Then, the following indicators were tested, including joint flexion of knee, leg circumference degrees, isokinetic muscle strength of the lower limb, feedback test, the test of YBT, the Balance Error Scoring System, feedforward test, static balance test on one foot, dynamic balance test on the feet, Lysholm knee score, anaerobic work of the upper extremity, torso strength, to analyze the graft healing, body shape, the knee joint function and physical quality improvement after ACL reconstruction. RESULTS AND CONCLUSION: After 10 months of phased rehabilitation training, the graft healed well; the knee flexion degree was back to normal; knee joint swelling basically eliminated; the muscle strength of the extensor flexor of the knee joint was back to over 95% of the normal level; the function of the injured knee joint was good and returned to a higher level; the Lysholm knee score was 85 points; the anaerobic work of the upper limbs and trunk strength reached to 120% of the normal level; and the quality of the body was greatly improved. To conclude, a great improvement in graft healing, knee joint function, recovery of muscle strength and physical qualities has been achieved in athletes who receive rehabilitation training after ACL reconstruction.

To get high cell density,pH-stat fermentations of L.casei Zhang were carried out in the medium optimized previously under different cultural conditions.The influences of neutralizing agents,the concen-tration of buffer salts and glucose in the medium,pH,aeration and fed batch fermentation on the growth of L.casei Zhang were investigated.Based on the values of maximum specific growth rate,biomass and viable cells count in different cultural conditions,the optimal growth condition was regarded as follows:Glucose level of the medium was 80 g/L ～100 g/L;The pH was kept constant at 5.9 with aqua ammonia and anae-robic condition was kept by sparged with nitrogen periodically;The temperature was set at 37?C and cul-tured for 10 h ～12 h.The biomass and viable cells' number of L.casei Zhang under this culture conditions were 7 g/L and 3.5?1010 CFU/mL respectively,and were 7 times as high as that before optimized,which can meet the requirements of probiotics.

Objective To investigate the association between Has‐miRNA‐149 polymorphism and susceptibility of digestive tract cancer .Methods A systematic search of Pubmed ,EMBASE and Web of Science ,CNKI ,Wanfang was conducted for relevant studies .The odds ratios(OR) and 95% confidence intervals(95% CI) calculated by Stata12 .0 software were used to assess the asso‐ciation between Has‐miRNA‐149 polymorphism and risk of digestive tract cancer .The Q test ,I2 value and funnel plot were conduc‐ted to examine heterogeneity and publication bias ,respectively .Results A total of thirteen eligible studies containing 4 424 cases and 5 290 controls were included in this Meta analysis .There were significant relationships between Hsa‐miRNA‐149 polymorphism and susceptibility of digestive tract cancer (Domina model CT+CC vs .TT ,OR=0 .915 ,95% CI:0 .840-0 .996 ,P=0 .040 ;Hetero‐zygous model CT vs .TT ,OR=0 .880 ,95% CI:0 .803 -0 .965 ,P=0 .007) .In the stratified analysis ,it was found that the risk of colorectal cancer decreased in patients carrying CC/CT genotypes(OR=0 .834 ,95% CI:0 .715-0 .972 ,P=0 .021) ,especially in A‐sians (CT+CC vs .TT ,OR=0 .894 ,95% CI:0 .818-0 .977 ,P=0 .013) ,while that was not found in the Caucasoids .Conclusion Has‐miRNA‐149 polymorphism is associated with the susceptibility of digestive tract cancer .The CC/CT genotypes may decrease the risk of digestive tract cancer ,especially in Asians .