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.

Objective To discuss appropriate strategy about treatment of rotatory fixation or dislocation of atlantoaxial articulation in children. Methods 32 cases of rotatory fixation or dislocation of atlantoaxial articulation in children were analyzed retrospectively from January 2000 to March 2005, 24 males and 8 females; patients' age was from 3 to 12 years old with an average of 5.5 years old. The mean course of diseases was 6.5 weeks (range: 2 days to 50 weeks). According to Fielding-Hawkins clinical classification, typeⅠ17 cases(1 case with os odontoideum), type Ⅱ 12 cases(1 case with os odontoideum and 1 case with congenital absence of the arch of posterior atlas), type Ⅲ 3 cases. 26 cases received conservational treatments which were recommended as 2-3 weeks of cervical traction, traction weight should be controlled to 1.0-1.5 kg. After a successful reduction a proper external fixation was required to maintain reduction. And another 6 serious cases received posterior atlantoaxial or occipitocervical autograft fusion. Indications for posterior atlantoaxial or occipitocervical atuograft fusion includes: 1) have difficulty in reduction even under proper traction; 2) with obvious neurological symptoms; 3) combined with compensatory deformity of atlantoccipital articulation or other occiptocervical deformities. Results All of 32 cases were followed up from 3 to 50 months with an average of 32 months. 26 cases who received cervical traction and external fixation resulted in satisfactory outcome in which all the torticollis were rectified, bilateral masses were symmetrical on AP position, and ADI was less than 4 mm in dynamic extension and flexation lateral view. ROM in rotation and extension and flexion were completely recovered. All the 6 cases surgically treated obtained sound bony fusion and neural symptoms were improved obviously and torticollis were rectified completely in 4 cases and others (2 cases) stopped progressing after operation. Conclusion Conservative treatment including Glission traction, cranial traction, collar, cast or cervicothoracic external fixation has been proved to be very effective in most of rotatory fixation or dislocation of atlantoaxial articulation in children; however, operative treatment should be considered in the following situations: patients with difficulty in reduction, with neural involvement and compensatory deformity of atlantoccipital articulation or other occipitocervical deformities.

[Objective] To study the methods of inducing bone mesenchymal stem cells(BMSCs)of rabbits into chondrocytes in vitro and the interaction of transforming growth factor ?1(TGF-?1),insulin-like growth factor-Ⅰ(IGF-Ⅰ)and basic fibroblast growth factor(bFGF).[Methods]BMSCs of rabbits were primarily cultured and subcultured in vitro,and then divided into four groups according to the difference of factors:group A receiving TGF-?1 and bFGF;group B receiving TGF-?1 and IGF-Ⅰ;group C receiving TGF-?1;group D receiving no cell growth factor.After three weeks all the four groups were detected by methyl thiazolyl tetrazolium(MTT)assay,measurement of glycosaminoglycan(GAG)and immunohistochemistry.[Results]Immunohistochemical detection of collagen Ⅱ was positive in groups A,B and C.The results of the MTT assay and the GAG content in groups A and B were obviously higher than those in groups C and D.[Conclusion]Rabbit BMSCs can be induced into chondrocytes under certain conditions.TGF-?1,IGF-Ⅰ and bFGF have synergy effect in the differentiation from BMSCs into chondrocytes.

Objective: To study the chemical constituents of the aerial parts of Siegesbeckia pubescens. Methods: A systematic separation of chemical constituents was conducted. Detailed chemical investigation of S. pubescens led to the isolation of six compounds by comprehensive chromatographic Methods: (Extraction, Silic gel C.C, Sephadex LH-20, ODS C. C). The structures of them were fully determined based on spectroscopic analysis including IR, HR-ESI-MS, ESI-MS, 1H-NMR, 13C-NMR, DEPT, HSQC, and HMBC spectrum. Results: A total of six compounds were isolated from the fraction of n-butanol extract of S. pubescens including one new monoterpenoid glycoside named (2Z,5E)-7-hydroxy-3,7-dimethyl-2,5-octadiene-1-O-[α-L-rhamnpyranosyl (1→6)]-β-D-glucopyranoside (1), four known diterpenoid glucosides named pubeside D (2), ent-15,16,19-trihydroxypimar- 8(14)-en-2-one-19-O-β-glucopyranoside (3), ent-15,16-dihydroxypimar-8(14)-en-2-one-19-oic-β-glucopyranoside (4), and neodarutoside (5), and eleutherazine B (6). Conclusion: Compound 1 is a new compound, named as siegeside F, and compound 6 is isolated from this species for the first time.

Objective To investigate the variation of serum Thymidine Kinase 1(TK1) in patients with non‐small cell lung canc‐er(NSCLC) and its clinical value .Methods Serum TK1 level of 129 patients with NSCLC and 90 healthy volunteers were measured by sensitive chemiluminescence dot‐blot assay .The serum TK1 level variation of 76 patients with NSCLC were compared before and after operation .Results Serum TK1 level of NSCLC patients were significantly higher than the level of healthy control group(P<0 .05) .Serum TK1 level in Stage Ⅲ >Stage Ⅳ >Stage Ⅰ + Ⅱ >Stage Tis ,comparison between each other was of statistical differ‐ence(P<0 .05) .The serum TK1 level of NSCLC patients at 30th day after surgery was remarkably lower than which before surger‐y(P=0 .001 ,P<0 .01) .The serum TKl level was correlated with lymphatic metastasis(P<0 .01) ,but not with other factors such as pathology types ,age ,sex and smoking .The serum TK1 measurement was high sensitivity and specificity to the diagnosis of NSCLC .Conclusion The serum TK1 level detection has important clinical significance in diagnosis and evaluation of NSCLC pa‐tients .

Objective To investigate the clinical value of the transluminal radiofrequency catheter ablation (RFCA) for malignant esophageal obstruction.Methods The retrospective cross-sectional descriptive study was conducted.The clinicopathological data of 52 patients with malignant esophageal obstruction who underwent transluminal RFCA at the Affiliated Hospital of Shandong Academy of Medical Science between March 2013 and March 2016 were collected.Patients received the bipolar radiofrequency ablation (RFA) under dualchannel endoscopy and X-ray.Observation indicators:(1) intra-and post-operative situations:operation situations,operation time,time of RFA,postoperative complications and duration of postoperative hospital stay,(2) follow-up.Follow-up using outpatient examination and telephone interview was performed to detect the subsequent treatment,survival of patients and recurrence of esophageal obstruction up to June 2016.Measurement data with normal distribution were represented as average (range).Results (1) Intra-and post-operative situations:52 patients underwent successful RFCA,without the occurrence of aspiration,asphyxia,hemorrhage and perforation.Esophageal obstruction was disappeared after treatment,X-ray findings showed a smooth esophagus.Average operation time and time of RFCA were respectively 58 minutes (range,20-71 minutes) and 23 minutes (range,8-42 minutes).Patients took liquid food at postoperative day 2 and normal food at postoperative day 3,without the sensations of esophageal obstruction.Of 52 patients,1 with postoperative hypotension returned to normal level through rehydration and increasing blood volume.Five patients with postoperative substernal pain were improved after 2-day symptomatic treatment.And other 46 patients didn't have postoperative complications.Average duration of postoperative hospital stay was 3 days (range,1-5 days).(2)Follow-up:52 patients were followed up for 3-24 months,with a median time of 13 months.Of 52 patients,17 underwent single intravascular interventional therapy,15 underwent intravascular interventional therapy combined with single systemic chemotherapy,14 underwent single systemic chemotherapy and other 6 didn't undergo antineoplastic therapy.During the follow-up,9 patients didn't have esophageal obstruction and 26 were complicated with esophageal obstruction again.Esophageal obstruction of 26 patients was respectively occurred at 3-8 months postoperatively,20 patients were improved after bipolar transluminal RFCA under dual-channel endoscopy and X-ray and 6 received parenteral nutrition support therapy due to extreme exhaustion.Seventeen patients died of cachexia caused by terminal malignant tumors.Conclusion Transluminal RFCA is safe and effective for malignant esophageal obstruction,with a good short-term outcome.

NSP4, as the diarrhea-related protein of rotavirus, is becoming an attractive candidate for vaccine development. To compare the immunogenicity of NSP4 from different genetic groups, we constructed eukaryotic expression plasmids comprising the NSP4 genes from four different genetic types using the pCI vector. The recombinant vectors were designated as pCI-97B6, pCI-97S36, pCI-97S34 and pCI-97SZ8, respectively. Following the conformation of the transient expression of the constructs in 293 cells, the plasmids were respectively subjected to the 5 round i. m. inoculation of BALB/c mice. The specific antibodies against NSP4 as well as the IgG1/IgG2a subclasses of immunoglobulin in mice sera were examined with indirect ELJSA after each immunization. The results showed that the immunization of plasmids expression NSP4s could elicit not only humoral but also cellular immunity, but the humoral immune response is dominant. There is a difference of immunogenecity among the NSP4 of different genetic type. Further studies were needed to focus on the relationship between the immunogenicity and protection effect.

ObjectiveTo investigate the targeting infection of single chain antibody againstAFP (scFv anti-AFP) directed lentivirus and the inhibitory effects of a dual-growth inhibition systemon hepatocarcinoma cells.MethodsPlasmids WtP53-pPRIME-miR30-shRNA-IGF1R,pMD2G-Anti-AFP,and psPAX2 have previously been constructed to cotransfect to the packaging cell line 293Tusing Lipofectamine2000.The infection results were observed through fluorescence microscopy.PCRand Western blotting were used to demonstrate the successful transduction and transcription of theWtP53-pPRIME-miR30-shRNA-IGF1R gene.The effects of reconstructed lentivirus infected liver cellgrowth were assessed by the cell growth curve of CCK8 cells. Apoptosis was evaluated by theTUNEL assay.ResultsRecombined lentivirus was successfully constructed with the functional PFUtiters of recombined lentivirus at 4.58× 109PFU/ml.This positive result was confirmed by PCR andWestern blotting.ConclusionsThe targeted therapy mediated by anti-AFP scFv could significantlyinhibit the proliferation of HEP3B cells and promote the apoptosis.

Adult ; Aged ; Aged, 80 and over ; Brachytherapy ; adverse effects ; methods ; Digestive System Neoplasms ; radiotherapy ; Esophageal Neoplasms ; radiotherapy ; Female ; Fever ; etiology ; Follow-Up Studies ; Humans ; Iodine Radioisotopes ; therapeutic use ; Male ; Middle Aged ; Nausea ; etiology

Objective To investigate the effects of alcohol on hippocampal apoptosis-inducing factor (AIF) and caspase-3 expression in cercbral ischemia/reperfusion in rats.Methods Sixty-eight healthy adult male Wistar rats were randomly divided into a sham operation group (n =4) and a saline control group as well as low-dose (1.0 g/kg),medium-dose (1.5 g/kg) and high-dose (2.0 g/kg) ethanol groups.The saline control group and each dose alcohol group were redivided into ischemia/reperfusion 0,1,2 and 3 h subgroups according to the intervention time points (n =4 in each group).A model of middle cerebral artery ischemia/reperfusion in rats was induced by the suture method.The corresponding doses of ethanol or an equal volume of saline were injected intraperitoneally at ischemia for 2 h and reperfusion for 0,1,2 and 3 h in the alcohol groups and the saline control group.At ischemia for 2 hand reperfusion for 24 h,the neurological deficit in rats was evaluated by using behavioral score.Immunohistochemistry assay was used to detect the expressions of AIF and caspase3 in the hippocampus of ischemic sides at ischemia for 2 hand reperfusion for 24 h.Results The behavior evaluation showed that the neurological deficit score was 0 in the sham operation group.The neurological deficit scores in the different dose ethanol groups were significantly lower than those in the saline control group at the same intervention time points (all P=0.000),and there were significant differences between the same intervention time points in the different dose ethanol groups (all P =0.000).The high-dose ethanol group was the lowest.There were no significant differences between the different intervention time points in the same dose ethanol groups (all P＞0.05).Immunohistochemistry revealed that the numbers of positive cells of AIF and caspase-3 in the sham operation group were 17.21 ±2.86 and 20.60 ±4.39,respectively,and they were significantly less than those in the saline control group and the each dose ethanol group at ischemia/reperfusion 0 h (all P =0.000); the number of positive cells of hippocampal AIF and caspase-3 in the different dose ethanol groups were all significantly less than those in the saline control group at the same intervention time points (all P =0.000).There were significant differences between the same intervention time points in the different dose ethanol groups (all P =0.000),and the high-does ethanol group was the lowest.There were no significant differences between the different intervention time points in the same dose ethanol groups (all P =0.000).Conclusions Alcohol has protective effect on the cerebral tissue in ischemia/reperfusion injury in rats.Its mechanism may be associated with the inhibition of AIF and caspase-3 expression.