Objective To compare the immune protective effects of three antigens of Trichinella spiralis encapsulated larvae on mice.Methods The mice were immunized with Trichinella spiralis encapsulated larvae somatic antigen,encapsulated larva excretory-secretory antigen and encapsulated larva surface antigen,3 times with a 7-day interval,and the adjuvant control and normal control group were set up.Seven days after the final immunization,each mouse was orally challenged with 200 Trichinella spirais larvae.The intestinal adult worms and muscle larvae of Trichinella spiralis of each group were recoveried and examined on Day 7 and Day 30 post-challenge,respectively.The level of 8eruln IgG to antigens of Trichinella muscle muscle larvae wa8 detected by ELISA.Results The intestinal adult worms were reduced by 84.89%.89.73%,85.65%.2.57% in the encapsulated larva somatic,excretory-secretory and surface antigen groups,respectively.The muscle lalwae were reduced by 71.71%,80.98%,73.66%,5.60%, respectively.Adtlltwornl reduction rates(P<0.05) and musclelarva reduction rates(P<0.01) of the encapsulated larva excretory-secretory antigen group and surface antigen group were higher than those of encapsulated larva somatic antigen group.The antibody titers in all the immunized groups increased significantly.and the GMRT values of the encapsulated larva somatic,excretory-secretory and surface antigen groups were 32 798.89,3 474.51,2 984.83,respectively,and were 6.09,7.56,6.50 times higher than those of the normal control group(459.32).Conclusions Trichinella spiralis encapsulated larva antigens can induce strong resistance of host to a subsequent challenge infection.Among these antigens,excretory-secretory antigen is more immunogenic.

Objective To investigate the association of human papilloma virus (HPV) typing and p53 expression with radiosensitivity in patients with cervical cancer.Methods A total of 80 patients with cervical cancer from 2014 to 2016 were enrolled,and among these patients,40 had stage Ⅰ B+ Ⅱ A disease and 40 had stage Ⅱ B +ⅢA disease.HPV genotype was identified and p53 expression was measured.All the patients underwent external and internal pelvic irradiation alone,and the correlation between short-term therapeutic effect and HPV typing/p53 expression was analyzed.The chi-square test or the Fisher's exact test was used for statistical analysis,and Spearman rank correlation analysis was also performed.Results The radiotherapy-insensitive (stable disease+progressive disease) group had higher p53 positive rates than the radiotherapy-sensitive (complete response+partial response) group (stage Ⅰ B+ Ⅱ:100％ vs.80.0％,P=0.044;stage Ⅱ B+ⅢA:100％ vs.90.0％,P=0.013).The expression of p53 was negatively correlated with radiosensitivity (r =-0.427,P =0.000).In the radiotherapy-insensitive group of patients with stage I B + Ⅱ and Ⅱ B +Ⅲ A,the rate of HPV multiple infections was higher than that of single subtype infection (65.0％/95.0％ vs.35.0％/5.0％,P=0.004 and 0.003),while in the radiotherapy-sensitive group,the rate of single subtype infection was higher than that of multiple infections (85.0％/60.0％ vs.15.0％/40.0％,P=0.004 and 0.003).The highest detection rate of HPV16 was 66.3％ in all patients,and the highest detection rate of HPV18 was 60.0％ in the radiotherapy-insensitive group.Conclusions High expression of p53 is associated with radioresistance in patients with cervical cancer.Patients with HPV muhiple infections have poor radiosensitivity,and HPV16 is the most common subtype in dual infection.Among patients who do not achieve remission after radiotherapy,HPV multiple infections with HPV18 as the main pathogen has the highest detection rate.

Objective: To observe influence of sensation of uncertainty in illness on curative effect of two-week rehabilitation program in patients with acute myocardial infarction (AMI). Methods: A total of 85 AMI patients recently treated in our hospital were selected continuously. They received two-week rehabilitation program treatment and were assessed by Missals uncertainty in illness scale (MUIS). According to MUIS scores, AMI patients were divided into middle-high score group (74.8~117.4 scores, n=51) and low score group (32~74.7 scores, n=34), and they were compared with 43 patients with coronary heart disease (CHD) angina pectoris (CHD control group) received treatment in the same period. Results: Compared with CHD control group, there were significant increase in each dimension score and total score of MUIS [total score (60.61±12.42) scores vs. (78.34±15.20) scores]in AMI patients (P<0.05~0.01). The exercise peak heart rate of MUIS middle-high score group was significantly higher than that of low score group [(137.80±26.49) times/min vs. (126.12±20.51) times/min, P<0.01]. Compared with low score group, there were significant decrease in total scores of activity of daily living (ADL) scale [(84.15±16.38) scores vs. (73.92±14.21) scores] and the 36-item short-form general heath survey [SF-36, (45.22±6.86) scores vs. (37.95±6.43) scores], and significant increase in total score of symptom checklist (SCL)-90 [(138.35±36.47) scores vs. (151.87±42.61) scores], mean days in CCU [(2.53±0.26)d vs. (2.77±0.29)d], mean days on bed [(4.46±0.25)d vs. (5.38±1.22)d], mean hospital day [(20.48±3.16)d vs. (25.37±3.82)d] and mean inpatient fee [(39.1±8.2) thousand RMB vs. (45.7±9.3) thousand RMB] in middle-high score group, P<0.05~0.01. Conclusion: There is significant sensation of uncertainty in illness in patients with acute myocardial infarction, and it makes curative effect of two-week rehabilitation program significantly decrease.

OBJECTIVE To compare different methods of processing of homograft costal cartilage. METHODS Homograft costal cartilage samples were processesd by three methods-irradiation, freezing, and alcohol fixation-before being inserted into the rabbit nasal dorsum. They were taken out 12 weeks later and evaluated for histological responses under a optical microscope. RESULTS The histological responses consisted of inflammatory cell infiltration, necrosis, vasal responses and fibroplasia. The was a significant different in degree of inflammatory cell infiltration and necrosis in all three groups(P

Objective To discover the reciprocal regulation and its molecular mechanism of estro-gen, IL-6 and IL-8 in ovarian cancer cells. Methods Based on our previous studies, the effect of 17β-estradiol (E2) on the expression levels of IL-6, IL-8 and their respective receptors was investigated. Mean-while, the effect of IL-6/IL-8 on estrogen receptor (ER) expression and estrogen-dependent transcriptional activation was analyzed. Gene expression profile analysis revealed that CAOV-3 and OVCAR-3 cells, which express ER, IL-6 and IL-8 receptors, were suitable models for this study. Results We found that E2 not only enhanced IL-6/IL-8 secretion via NF-κB signaling pathway, but also modulated IL-6 and IL-8 receptors expression. Tamoxifen (Txf), an ER antagonist, completely abolished E2-stimulated IL-6/IL-8 expression. On the other hand, in the absence of estrogen, both cytokines increased ERα expression, decreased ERβ ex-pression, and activated estrogen-dependent transcriptional activation, which was completely blocked by Txf. Pretreatment of OVCAR-3 with p38 MAPK, MEK1/2 or ErbB2 MAPK inhihitors, respectively, IL-6-media-ted ER activation was blocked, while IL-8-indueed ER activation was blocked by Src inhibitor. Conclusion These data suggest that estrogen, IL-6 and IL-8 may form a mutual amplifying signaling which contributes to the growth and development of ovarian carcinoma.

Objective To observe the mitochondrial damage associated with protein-energy wasting of skeletal muscle in diabetic kidney disease (DKD) model of Goto-Kakizaki(GK) rats and evaluate the effects of low-protein diet supplemented with α-keto acids on muscle wasting.Methods Forty-five male 24-week-age GK rats were randomly divided into three groups,normal protein diet group (NPD),low-protein diet group (LPD) and LPD +or-keto group (Keto).Fifteen gender and age matched Wistar rats were served as control group (CTL).The living condition of GK rats was observed and the weight was measured once a week.Urine albumin,serum glucose,creatinine and urea nitrogen were measured at 24,32,40,48 week age.Soleus muscle was observed to calculate the muscle size and the percentage of Ⅰ and Ⅱ type muscle fiber with software after SDH and NADH staining at 48-week-age.Tissue ultrastructure was observed under the transmission electron microscopy.The activity of citrate synthase was detected by spectrophotometer.Expression of mitochondrial DNA was examined by Q-PCR.Results Compared with the CTL group,NPD,LPD and Keto groups had lower body weight,higher urine albumin,higher serum creatinine and urea nitrogen (P ＜ 0.05).The crosssectional area of muscle fibers was larger in CTL group.Compared with CTL group,the muscle fiber was partly broken,the mitochondrial morphology was obviously changed,the percentage of type Ⅱmuscle fiber was increased significantly (P ＜ 0.05),and the activity of citrate synthase and the number of mitochondrial DNA were decreased significantly in NPD,LPD and Keto groups (P ＜ 0.05).In Keto group,muscle wasting was improved compared with NPD and LPD group (P ＜ 0.05),the crosssectional area of soleus muscle increased and the percentage of type Ⅱ muscle fiber decreased,levels of urine albumin,semm creatinine and urea nitrogen decreased (P ＜ 0.05).Under transmission electron microscopy,the muscle fiber of keto group was intact and mitochondiral morphology was close to that of CTL group.The activity of citrate synthase and number of mitochondiral DNA were higher as compared to CTL group (P ＜ 0.05).There were no significant differences between NPD and LPD group.Conclusions In DKD condition,protein degradation in the skeletal muscle is accelerated,mitochondrion is swelling,the number of mitochondrial DNA is decreased and mitochondrial function is impaired.Low-protein diet supplemented with α-keto acids can improve mitochondrial damage and muscle wasting induced by DKD.

Objective To investigate the characteristics of co-infection in initial treatment acute leukemia induction chemotherapy.Methods The clinical features of 179 untreated acute leukemia patients with nosocomial infection were analyzed after combined chemotherapy.Results In the 179 patients,151cases achieved complete remission,the complete remission rate was 84.4 ％,82 cases suffered from nosocomial infections,the incidence of infection was 45.8 ％.The sites of infection were oral,anal,lung,as well as primary foci was not clear bacteremia.In 428 specimens,the isolated bacterial colony counted a total of 66,the number of fungal colonies was 9,the bacterial colony was G-bacteria-based.G-bacteria had different degrees of resistance to many antibiotics.Extended-spectrum β-lactamases strains had not been detected in these specimens.Conclusion Acute leukemia patients is easy to co-infection after chemotherapy.Control and prevention of nosocomial infections should run throughout the entire treatment process,application of laminar flow bed helps reduce the newly diagnosed acute leukemia patients with nosocomial infection incidence.

Background The application of mesenchymal stem cells to transfer specific genes is under investigation in various diseases.Using this strategy may provide a more effective method to supply exogenous neurotrophic factors to the cental nervous system,including retina.Objective This study was to construct ciliary neurotrophic factor(CNTF)-overexpressing bone marrow mesenchymal stem cells (BMSCs) using lentiviral vectors.Methods Rat secreted-CNTF gene cDNA was synthesized and subcloned into a lentiviral vector plasmid pHⅣ-dTomato to construct recombinant vector CNTF-dTomato.CNTF-dTomato/pH Ⅳ-dTomato plasmid were co-transfected into 293T packaging cell line with packaging plasmid psPAX2 and enveloped plasmid pMD.2G to produce recombinant lentivirus CNTF-lenti and control-lenti.Rat BMSCs were infected with CNTF-lenti/control-lenti to produce CNTF-BMSCs and control-BMSCs.Expression of dTomato and efficiency of infection was evaluated under the fluorescence microscope.Uninfected BMSCs(pure BMSCs) served as the blank control.CNTF protein level in the supernate was detected by enzyme-linked immunosorbent assay (ELISA) and compared among the blank-BMSCs group,control-BMSCs group and CNTF-BMSCs group.Adipogenic and osteogenic differentiation of CNTF-BMSCs were induced using adipogenic-inducible medium and osteogenic-inducible medium and identified using oil-red O staining and alizarin red S (ARS) staining.Results After CNTF-dTomato plasmid was transfected into Stbl3 competent cells,the colony PCR product was 1 033 bp.The inserted sequence in the pHⅣ-dTomato plasmid was coincident with the expected one.The results of DNA sequencing showed that CNTF-dTomato plasmid was successfully constructed.The infection rate of CNTF-lenti was approximately 95％.ELISA showed that on the post-infected day 2,3,7,the CNTF protein levels in the supernate were significantly higher in the CNTF-BMSCs group than those in the blankBMSCs group and control-BMSCs group (all at P=0.000).In the CNTF-BMSCs group,the CNTF protein levels in the supernate were significantly increased on post-infected day 2,3,7 compared with day 1 (P =0.013,0.004,0.042).The adipogenic-induced cells showed the red staining to oil red O,and osteogenic-induced cells presented the orange staining to ARS.Conclusions BMSC line with stable expression of CNTF is successfully established by lentiviral vectors.CNTF-BMSCs have the potential to differentiate towards adipocytes and osteoblasts.

Background Krüppel-like factor 6 (KLF6) is related to the physiological or pathological process,such as growth,cell differentiation,proliferation,apoptosis,angiogenesis,tissue repair,and so on.But in ophthalmology,it is less reported about the expressing level of KLF6 protein in lens epithelial cclls (LECs) or the effect of KLF6 on the proliferation of human LECs.Objective This study was to investigate whether KLF6 can inhibit proliferation of human LECs.Methods KLF6 eukaryotic expression plasmid (pEGFP-C2-KLF6) was constructed using reverse transcription PCR(RT-PCR) and identified by double enzyme digestion method and PCR.Human LECs strain (HLE-B3) was cultured and passaged using low glucose DMEM containing 10％ fetal bovine serum and then divided into 4 groups.KLE-B3 transfection reagents were added in the culture medium of all groups.In addition,no agent was used in the blank control group;only insulin-like growth factor-1 (IGF-1) was appended to the medium in only IGF-1 group ;null vector was transfected and IGF-1 was appended in the null plasmid transfection+ IGF-1 group;while pEGFP-C2-KLF6 eukaryotic expression plasmid was transfected into the cells,and simultaneously IGF-1 was added in the pEGFP-C2-KLF6 plasmid transfection+IGF-1 group.After 24 hours of intervene,water soluble tetrazolium salt-1 (WST-1) test was used to detect the growth status of the cells,and Western blot assay was used to assay the relative expressing level of KLF6 protein in the cells.In the other hand,the cells were cultured at the density of 1 ×104/piece,and 0,0.10 and 0.25 μg pEGFP-KLF6 were transfected into each piece of cells respectively,and then IGF-1 was added with a final concentration of 50 μg/L for 24 hours after cell culture.Expressions of Ki-67 protein and mRNA in the cell pieces were detected by immunocytochemistry and fluorescent semiquantitative PCR,respectively.Results The PCR product bands were consistent with KLF-6 gene in length,and the product fragments were corresponding with expectant ones via PCR and double enzyme digestion method,showing a successful construction of pEGFP-C2-KLF6 eukaryotic expression plasmid.After 24 hours of IGF-1 stimulation,the absorbance values of the cells were 0.86±0.00,2.10±0.01,2.24±0.12 and 1.06±0.02 in the blank control group,only IGF-1 group,null plasmid transfection + IGF-1 group and the pEGFP-C2-KLF6 plasmid transfection + IGF-1 group,with a significant difference among the 4 groups (F =38.322,P ＜ 0.05),and that in the pEGFP-C2-KLF6 plasmid transfection +IGF-1 group was significantly lowed in comparison with the only IGF-1 group and the null plasmid transfection+IGF-1 group (q=6.42,7.31,both at P＜0.05).Western blot assay showed that the relative expressing levels of KLF6 protein were statistically different among the four groups (F =591.858,P＜0.05),and those in the pEGFP-C2-KLF6 plasmid transfection+IGF-1 group were 1.47,2.04,3.27 folds higher than those in the blank control group,only IGF-1 group,respectively and null plssmid transfection+IGF-1 group,respectively.Immunocytochemistry revealed that the expressing intensity of Ki-67 protein was gradually weakened with the decrease of pEGFP-C2-KLF6 plasmid dose in the 0,0.10 and 0.25 μg pEGFP-C2-KLF6 plasmid transfection+IGF-1 groups.Fluorescence semiquantitative PCR results showed that the relative expression values of Ki-67 mRNA in the cells were O.15±0.08 and 0.11±0.03 in the 0.10 μg and 0.25 μg pEGFP-C2-KLF6 plasmid transfection+ IGF-1 groups,which were significantly lower than O.77± 0.12 of the 0 μg pEGFP-C2-KLF6 plasmid transfection group,with a statistically significant difference among the three groups (F =54.825,P＜0.05).Conclusions KLF-6 can effectively inhibit IGF-1-induced proliferation of human LECs,and it can be regarded as one of the regulatory factors of the proliferation of HLE-B3 cells.

Pi.Au-Pt alloy does not resist the growth and adhesion of the bacteria, but Au-Pd does.