Objective To construct lentiviral vector miR30-CD133 targeting CD133 gene and investigate its effects on SMMC7721 cells. Methods Thespecific sequence of DNA which containing both sense and antisense Oligo DNA of the targeting sequence were designed, synthesized and cloned into the pPRIME vector. pPRIME-miR30-CD133, psPAX2 and pMD2G were co-transfected on 293T cells to get the lentivirus the transfection efficiency was investigated by confocal laser scanning microscope. The expression of miR30-CD133 on CD133 were detected by qRT-PCR and Western blott. The proliferation and apoptosis effect of miR30-CD133 was respectively evaluated by CCK-8 assay and AnnexinV/PI. Results Functional PFU titers of PRIME-miR30-CD133 were 6.58 × 109 PFU/mL. The expression of CD133 mRNA in sh CD133 group (0.18 ± 0.04) was less than Blank group and shNC group (P<0.05). The expression of CD133 protein in sh CD133 group was less than Blank group (10%) and shNC group (8%) (P < 0.05). Cells proliferation activity were significantly inhibited in shCD133 group compared with control group. Apoptosis rate were significantly increased in shCD133 group (41.3%) compared with Blank group (25.3%)and shNC group (24.3%). Conclusion The lentivirus miR30 vector targeting CD133 gene was successfully constructed, which will be a basis to the further CD133 gene studies.

ObjectiveTo observe the effect of family rehabilitation plan (FRP) on motor function of stroke patients with hemiplegia.Methods64 patients were randomly divided into the FRP group (32 cases) and control group (32 cases). The patients in the FRP group critically followed the schedule and activities scheduled on FRP and were assessed and guided in out-patient department every two weeks. The patients in the control group made schedule and carried on activities by themselves. Motor function was assessed with Fugl-Meyer Assessment (FMA) and modified Barthel index (MBI) respectively.ResultsMotor scores of the patients in two groups significantly increased after treatment ( P<0.001), and that of the FRP group was significantly better than that of the control group ( P<0.001).ConclusionThe FRP can significantly improve motor function of stroke patients with hemiplegia.

To detect the expression of p16 protein and Ki-67 antigen in gastric mucosa-associated lymphoid tissue lymphoma(MALTL)and normal gastric mucosa and to investigate the clinical significance of their expression in the occurrence and development of gastric MALTL.Methods 47 samples of gastric MALTL diagnosed pathologically in the department of pathology of the PLA General Hospital from March 1993 to June 2005 were collected.By using immunohistochemial methods,the expression of p16 protein and Ki-67 antigen was detected in 20 samples of normal gastric tissue and the 47 samples of gastric MALTL.Results The positive rate of p16 protein Was 21.3%(10/47)and 90.0% in gastric MALTL and normal gastric tissue respectively.The positive rate of p16 protein in gastric MALTL was lower than that in normal gastric tissue(P<0.05).The expression of p16 was related to the degree of lymph node metastasis.The positive rate of Ki-67 labelling index(LI)in gnstric MALTL was hiigher than that in normal gastric tissue.A negative correlation was found between the expression of p16 protein and Ki-67 LI(P<0.05). Conclusions Detection p16 and Ki-67 may help to predict the possibility of lymph node metastasis and prognosis in gastric MALTL.

Object To study the chemical constituents with the antiviral effect from Yinqiaosan on influenza virus Methods Isolation of the constituents by different kinds of column chromatography and elucidation of their structures on the basis of chemical and spectral methods Results Four flavonoid glycosides were isolated and elucidated as 2′ O D apiofuranosyl (1→2) ? D glucopyranosyl] isoliquiritigenin, 4′ isoliquiritigenin, 4′ O D apiofuranosyl (1→2) ? D glucopyranosyl] isoliquiritigenin, 4 isoliquiritigenin, 4 O D apiofuranosyl (1→2) ? D glucopyranosyl] isoliquiritigenin and 4′ isoliquiritigenin and 4′ O D apiofuranosyl (1→2) ? D glucopyranosyl] liquiritigenin, respectively liquiritigenin, respectively Conclusion 2′ O D apiofuranosyl (1→2) ? D glucopyranosyl] isoliquiritigenin is a new naturally compound isoliquiritigenin is a new naturally compound

Objective To investigate the gastrointestinal symptoms in patients with chronic obstructive pulmonary disease (COPD) and explore its possible reasons.Methods Thirty-six patients with stable COPD were divided into mild group [21 cases,forced expiratory volume in first second (FEV1) measured values/predictive values ≥50％] and severe group (15 cases,FEV1 measured values/predictive values ＜ 50％) according to the pulmonary function.Replacement of 19 healthy volunteers as control group.The patients and volunteers completed the Gastrointestinal Symptom Rating Scales (GSRS) scores and received electrogastrography,the plasma vasoactive intestinal peptide (VIP) and intestinal fatty acid binding protein (Ⅰ-FABP) were detected.Results The GSRS scores in severe group and mild group was higher than that in control group[(17.73 ± 4.64),(14.29 ± 5.44) scores vs.(8.00 ± 2.29) scores],and severe group was higher than that in mild group,there was significant difference (P ＜ 0.05).The percentage of normal slow waves of fasting and postprandial,the percentage of bradygastria in severe group and mild group were lower than those in control group,there were significant differences(P ＜ 0.05).The level of VIP in severe group was lower than that in mild group and control group [(36.07 ± 9.22) μ g/L vs.(44.16 ± 7.88),(46.53 ± 4.54) μ g/L],the level of I-FABP was higher than that in mild group and control group [(35.80 ± 11.69) μ g/L vs.(27.40 ±9.53),(23.58 ±8.18) μg/L],there was significant difference (P ＜0.05),but there was no significant difference between mild group and control group (P ＞ 0.05).Conclusion The patients with COPD are accompanied with serious gastrointestinal symptoms and gastric dysrhythmia,severe COPD can be associated with VIP decrease and Ⅰ-FABP rise in peripheral blood.

Objective To evaluate the predictive value of plasma atrial natriuretic peptide (ANP) level on prognostic of patients with systemic inflammatory response syndrome by dynamic monitoring ANP levels.Methods Ninety-eight patients admitted to the intensive care unite were classified into survival group(n =78) and death group (n =20).The level of plasma ANP,procalcitonin,C-reactive protein and lactic acid were measured.Acute Physiology and Chronic Health Evaluation (APACHE) Ⅱ score were recorded.Results The plasma ANP level of patients in the death group was 0.40 (0.26) μg/L,significantly higher than that in the survival group(0.22(0.12) μg/L,P =0.000).Along with treatment scheme,the plasma ANP level decreased in survival group,and there were significant difference among three times (0.22 (0.12) μg/L,0.17 (0.09) μg/L,0.13 (0.11) μg/L,P =0.000).But there was no difference in ANP level of patients in death group along with the disease developing (0.38 (0.30) μg/L,0.39 (0.23) μg/L,0.39 (0.22) μg/L,P =0.99).ICU hospitalized time in survive group associated with APACHE Ⅱ score,ANP and PCT(r =0.735,0.628,0.487 respectively,P =0.000,0.001,0.021).Conclusion ANP is proved to be a good clinical index in prognostic evaluation of patients with sepsis.

Objective To evaluate the regulatory effect of OX40 co-stimulatory signal on the expression of Foxp3 in inductive regulatory T cells (iTreg) in vitro.Method CD4+ CD25+ naive T cells were isolated from C57BL/6 mouse lymphocyte suspension by MASC CD4+ CD25+ regulatory T cell isolation kit.Inductive Tregs were generated by stimulation of naive T cells in the presence of transforming growth factor beta (TGFβ1),anti-CD3,anti-CD28 and IL-2.The regulatory effect on iTregs was shown by use of OX40 stimulation monoclonal antibody (OX86) or control antibody.Using flow cytometric analysis (FACS),we examined the antibody-based identification of Tregs surface markers CD4 and CD25,along with the intracellular activation marker FoxP3.Results The ratio of CD4+ CD25+ nTregs isolated from mouse lymphatic node was (5.0 ± 0.4)％ vs.(71.8 ± 13.4)％ of TGFβ1-driven iTregs.The ratio of CD4+ CD25+ Tregs was (80.0 ± 1.6) ％ in OX40 stimulation McAb group vs.(86.0 ± 1.4)％ in control antibody group.Furthermore,the expression of Foxp3 was (59.2 ± 0.7) ％ in OX40 stimulation McAb group vs.(70.0 ± 0.8) ％ in control antibody group (P＜0.05).Conclusion TGFβ1-dependent protocol may induce the conversion of naive CD4+ T cells into CD25+ Foxp3+ iTregs.OX40 stimulation can down-regulate the expression of Foxp3 in CD4+ CD25 + iTreg significantly.Thus OX40 molecular may become an attractive target in Tregs-induced transplant tolerance.Further study should be performed to increase the suppressive activity of iTregs through blockade of OX40 signal.

AIM: To evaluate the clinical outcomes and evaluate the application value of pedicle screw system internal fixation in short segment for the treatment of Hangman's fracture. METHODS: A retrospectively analysis was conducted in 17 cases of Hangman's fracture who were treated in Department of Orthopaedics, Sichuan Provincial People's Hospital since 2004. According to Levine-Edwards classification, 4 of them belonged to type II, 6 type IIA, 7 type III. According to Frankel classification in function of spinal cord, 12 cases of them belonged to Grade E and 5 cases D. All cases were fixed with C2,3 pedicle screw system short segment implantation materials (product of Medtronic Sofamor) after skull traction and reduction. RESULTS: No vertebra artery injury or spinal injury was found during operation. Postoperational photograph and CT detection revealed bilateral fracture had no reduction in 1 case and cerebrospinal fluid leaking occurred in 1 case who was cured after conventional therapy. All cases were followed up for 12-24 months (average 14.5 moths). All fractures healed within 6 months, and activities of cervical vertebra were not obviously limited. As for nerve function, 4 cases of Grade D recovered up to Grade E. Ti-alloy implant materials used in this study were well biocompatible to the human body. There were no blood, immune, histology reactions, or the corrosion and abrasion of Ti-alloy appeared. CONCLUSION: Pedicle screw fixation in short segment for the treatment of Hangman's fracture is satisfactory in fracture reduction, fixation and healing. There are little lose in function of cervical vertebrae and it promotes functional recovery of spinal cord.

Objective To investigate the efficacy and safety of dipeptidyl peptidase 4(DDP-4)inhibitor in the treatment of diabetes mellitus.Methods Eighty-six cases patients with poor glycemic control in type 2 diabetes mellitus in Shunde First People's Hospital of Foshan from May 2015 to May 2016 were selected as the research objects and divided into two groups according to random number table method,each group with 43 cases.The control group with acarbose(50 mg/times,3 times/d,with the same as the 3 meals,sustained medication for 12 weeks)to control blood sugar,while the observation group used DPP-4 inhibitors sitagliptin(100 mg/times,1 times/d,sustained medication for 12 weeks),compared the blood glucose,blood lipid indexes and adverse reactions of two groups before and after treatment.Results After treatment,FPG,2 hPG,HbAlc level of observation group were respectively(6.71±0.65)mmol/L,(8.10±0.17)mmol/L,(7.12±0.41)％,significantly lower than control group((7.86±0.72)mmol/L,(9.20±0.65)mmol/L,(7.51±0.52)％,the differences were statistically significant(P＜0.01).After treatment,there were no significant differences in terms of TC,TG,HDL-C,LDL-C between two groups(P＞0.05).Treatment for 12 weeks,the blood glucose compliance rate of observation group was 88.37％(38/43),the control group was 67.44％(29/43),the observation group was significantly higher than the control group(P=0.019).After treatment,the HOMA-IR value of the observation group was 4.42±0.17,significantly lower than the control group(4.91±0.24),HOMA-beta value was 88.20±6.31,significantly higher than that of the control group(80.21±5.67),the differences were significant(P＜0.01).Conclusion DDP-4 inhibitor in the treatment of type 2 diabetes,can effectively reduce the level of blood glucose and glycosylated hemoglobin,and have no significant adverse reactions,is effective and safe hypoglycemic drugs.

Objective To evaluate the therapeutic effect of esophageal intraluminal stents with 125I seeds based on survival time and quality of life (QOL) of patients with advanced esophageal cancer.Methods A total of 37 patients with advanced esophageal cancer who underwent placement of esophageal 125I irradiation stent were followed up.The QOL was evaluated by QLQ-C30 and QLQ-OES18 questionnaires before the procedure, and 1 week and 3 months after.Results The mean survival time of 37 patients was 28 weeks, with 3-month and 6-month survival rates at 94.6% (35/37) and 51.4% (19/37), respectively.Compared with the baseline, the scores of emotional functioning(EF), cognitive functioning(CF),global quality of life(QL), fatigue(FA), pain(PA), financial difficulties(FI) and pain of QLQ-OES18(OESPA) assessed 1 week after stent placement increased (P<0.05) significantly and dysphapia of QLQ-OES18(OESDYS), eating of QLQ-OES18(OESEAT), trouble swallowing saliva of QLQ-OES18(OESSV) and choked when swallowing of QLQ-OES18(OESCH) decreased significantly (P<0.05).At 3 months after stent placement, the scores of FI increased significantly (P<0.05), and those of nausea or vomiting (NV), OESDYS,OESEAT, OESSV and OESCH decreased significantly (P<0.05).The scores of FI,OESDYS, OESCH, role functioning(RF), EF,CF,FA and OESPA at 3 months were significantly different from those at 1 week after the procedure (P<0.05).Other variables at 1 week and 3 months after the procedure were not different from those before (P>0.05).Conclusion Implantation of 125I seeds combined with esophageal intraluminal stents may prolong survival time and improve QOL of patients with advanced esophageal cancer.