BACKGROUND:Total flavonoids of hippophae have different degrees of regulation effect on the immune system. However, there is no available evidence addressing the effect of total flavonoids of hippophae on vascular dementia inflammatory factor. OBJECTIVE:To observe the effect of total flavonoids of hippophae on serum tumor necrosis factor-αand interleukin-1βlevels in a rat model of vascular dementia, and to discuss its interventional mechanism for vascular dementia. METHODS:A total of 72 Wistar rats were randomly divided into six groups:sham operation, model, control, high-dose, medium-dose, and low-dose of total flavonoids of hippophae groups. Except for the sham operation group, rats in the other five groups were applied to establish vascular dementia models using the improved“vascular occlusion”method. Sham operation group was only subjected to the isolation of bilateral common carotid artery, without ligation. At 1 day after modeling, rats in control group was intragastrical y given Ginkgo biloba extracts 48 mg/kg?d, while rats in total flavonoids of hippophae groups were given 200, 100, 50 mg/kg?d of total flavonoids of hippophae. Model group and sham operation group received equal volume of saline. After 30 days of administration, rats were detected in Morris water maze. Serum tumor necrosis factor-αand interleukin-1βlevels of rats in different groups were detected after intervention. RESULTS AND CONCLUSION:Compared with sham operation group, the vascular dementia model rats had a disordered learning and memory ability, and increased levels of serum tumor necrosis factor-αand interleukin-1β(P<0.05). After intervention of Ginkgo biloba extracts and total flavonoids of hippophae, the learning and memory abilities were improved, and serum tumor necrosis factor-αand interleukin-1βlevels were decreased (P<0.05). Total flavonoids of hippophae can reduce serum tumor necrosis factor-αand interleukin-1βlevels in vascular dementia rats. This may be the main mechanism of treating vascular dementia.

Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a (+) vector, then ICP27 protein was expressed in E. Coli and purified by nickel-nitrilotriacetic acid (Ni2+-NTA) affinity resin column,finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay (IFA). Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27's biological function during HSV-1 infection.

Objective: To search for the optimizing parameters and distribution pattern of NF ?B decoy oligodeoxynucleotides in transfecting J774.1 cells mediated by lipofectin. Methods: With the change of ODNs/lipofectin ratio and transfection time, the uptake rate and mean fluorescence intensity of NF ?B decoy oligodeoxynucleotides in J774.1 cells were measured by flow cytometry to evaluate transfection efficiencies. Intracellular distribution of NF ?B decoy oligodeoxynucleotides was determined with fluorescence microscopy. The lactate dehydrogenase (LDH) activity in the supernatant was assayed to assess the cytotoxicity. Results: The uptake of NF ?B decoy oligodeoxynucleotides into J774.1 cells was significantly improved by lipofectin. In 24 well culture plate, 1 ?g ODNs with 5?g lipofectin ( W/W =1∶5) resulted in the highest transfection efficiency and lower cytotoxicity. The NF ?B decoy oligodeoxynucleotides localized in the nucleus as well as in the cytoplasm following an incubation of 6 h with lipofectin. While NF ?B decoy oligodeoxynucleotides had faint fluorescences in cytoplasm in the absence of lipofectin. Conclusion: lipofectin can enhance the cellular uptake of NF ?B decoy oligodeoxynucleotides in J774.1 cells and alter intracellular distribution of NF ?B decoy oligodeoxynucleotides. Efficiency of transfection is the highest when the ratio of ODNs/lipofectin is 1∶5 for incubation of 6 h.

Objective To investigate the safety,feasibility and effectivity of laparoscopic left hemihepatectomy for left hepatolithiasis.Methods From Jan.2005 to Dec.2013,36 patients with left hepatolithiasis underwent laparoscopic left hemihepatectomy (group LH),in comparison with 39 other patients who underwent conventional open left hemihepatectomy (group OH).The blood supply to left liver was dissected and cut off first.The liver parenchyma was transected and the left hepatic vein was dissected and clamped.Intraoperative choledochoscopy was carried out through the stump of left bile duct,laparoscopic choledocholithotomy and T-tube drainage were carried out when stones were found in the common or right bile duct.Blood loss,rate of residual stone,complication rate between the two groups were compared.Results The success rate of operation was 100％.Compared with group OH,group LH had shorter postoperative hospitalization,less incision infection and fewer analgesia needed(t =3.75,x2 =4.11,x2 =22.12,P ＜0.05).There was no statistical difference for blood loss,rate of transfusion,and postoperative complications such as bile leakage,pleural effusion,ascites,residual stones (t =0.66,x2 =0.70,x2 =0.01,x2 =0.52,x2 =0.01,x2 =0.28,x2 =0.01,P ＞ 0.05).Conclusions Laparoscopic left hemihepatectomy is safe,feasible,and effective for hepatolithiasis of the left liver lobe.

AIM: To study the effects of nuclear factor-kappa B decoy oligodeoxynucleotides on generation of nitric oxide and activated oxygen in lipopolysaccharide-induced mouse macrophages.METHODS: Mouse macrophage cell lines J774.1 cells were cultured with lipopolysaccharide and liposome-mediated oligodeoxynucleotides, and the level of nitric oxide, activated oxygen and inducible nitric oxide synthase were measured. RESULTS: Nuclear factor-kappa B “decoy”oligodeoxynucleotides decreased the activity of inducible nitric oxide synthase in lipopolysaccharide-induced macrophage, inhibited generation of nitric oxide and reactive oxygen species. The level of nitric oxide, activated oxygen and inducible nitric oxide synthase did not change in control group.CONCLUSION: Nuclear factor-kappa B “decoy”oligodeoxynucleotides decrease the generation of nitric oxide and activated oxygen in lipopolysaccharide-induced macrophages, which is probably due to inhibiting the binding sites of activated nuclear factor-kappa B specially.

AIM: To study the effect of NF-?B "decoy" oligodeoxynucleotides on TNF-? and IL-6 expression in LPS-induced mouse macrophages. METHODS: Mouse macrophage cell line J774.1 cells were cultured with LPS and liposome-mediated oligodeoxynucleotides, and the levels of TNF-? and IL-6 measured in the different culture supernatant by enzyme linked immunosorbent assay. RNA was extracted from macrophages, and the mRNA expression of TNF-? and IL-6 in macrophages was observed by RT-PCR. RESULTS: NF-?B "decoy" oligodeoxynucleotides decreased the expression of TNF-? and IL-6 in LPS-induced macrophages and inhibited generation of TNF-? and IL-6. The level of TNF-? and IL-6 did not change in control group. CONCLUSIONS: NF-?B "decoy" oligodeoxynucleotides inhibit the expression of TNF-? and IL-6 in LPS-induced macrophages, which is probably due to the specific inhibition of activated NF-?B binding sites .

Abstract Recent years have witnessed more attention paid to the health effects of the natural radionuclide 226Ra, and a series of research carried out on the detection methods of 226Ra at home and abroad. The common 226Ra detection methods are emanation scintillation, gamma spectrometry, liquid scintillation counting, alpha spectrometry and alpha counting. Emanation scintillation is mature enough as a classical method, while gamma spectrometry and liquid scintillation counting proceed fast. Due to the radiation of 226Ra, these detection methods still have the problems such as the requirement for large samples, long detection periods, complicated steps, high detection limits and low recovery rates. This paper reviews the five methods above and summarize their advantages and disadvantages, so as to provide the reference for selecting and optimizing the detection methods for different samples as well as perfecting the standard method in China.

Objective To evaluate the therapeutic efficacy of stereotactic body radiotherapy (SBRT) with gamma knife on stage Ⅰ-Ⅱ non-small-cell lung cancer(NSCLC)and the quality of life of the patients undergoing this therapy.Methods Twenty NSCLC patients with the median age of 76,10 at stage Ⅰ and 10 at stage Ⅱ who were unable or unwilling to undergo surgery were given SBRT with gamma knife at the doses of 3-6 Gy in 8-15 fractions,finished within 2 to 3 weeks.The prescription isodose line was 50％,the marginal dose was 39-56 Gy,the central dose was 78-112 Gy,and the total biologically effective dose was 51-83 Gy.The patients were observed after admission and followed up by chest CT 1,3,6,and 12 months after treatment until progressive disease or death.EORTC QLQ-LC43 questionnaire was used to investigate the changes in quality of life.Results The 20 patients were followed up for 24 (12-46) months.At six months after the treatment,the overall response rate was 80％,and the complete response rate was 35％.The 1,2 and 3-year local control rates were 100％,95％ and 95％,respectively.The 1,2 and 3-year overall survival rates were 95％,80％ and 50％ respectively; The 1,2,and 3-year progression free survival rates were 85％,64％ and 33％,respectively.The failure rate was 20％ and the rate of progress within the planning target volume was 5％.No acute toxicity at grade 3 and over occurred in any patient during the treatment.15％ of the patients developed grade 1-2 radiation pneumonia.Age,gender,pathologic index or not were weakly correlated with the overall survival.The emotional function was improved significantly after treatment (P ＜ 0.05),dyspnea and cough were improved at different degrees,however,not significantly.There were no significant changes in the physical function and symptoms,such as fatigue,lack of appetite,insomnia,etc.Conclusions Significantly improving the motional function and maintaining the quality of life,SBRT with gamma knife is effective for elderly NSCLC patients with high local control rate fair overall survival rate and few side effects.

Objective To evaluate the efficacy of stereotactic body radiation therapy (SBRT) for hepatic metastases from colorectal cancer,and to collect data for the application of this technique.Methods A total of 28 patients from No.306 Hospital of PLA,including 17 male and 11 female with median age of 63.8 (range from 31 to 86),were treated with SBRT for colorectal cancer with liver metastases with 54 lesions in total.The GTV,CTV and PTV were delineated above the enhanced CT scans acquired during normal quiet respiration.CTV was obtained by adding 5 mm isotropic margin from GTV,and PTV was obtained by adding 5 to 10 mm isotropic margin from CTV.Prescription dose line covered 50％-60％ of isodose curve at 3-6 Gy/fraction.The total dose was 39-45 Gy and the biologically equivalent doses(BED)was 50.7-65.3 Gy.The patients were followed-up beginning at 3 months after SBRT.The change in size of the lesion based on enhanced CT or MR scans was evaluated.Toxicity was evaluated and scored according to the RTOG criteria.Local control rate and survival rate were analysed.Results All patients completed the treatment.With median follow-up of 15.1 months (range frome 3 to 30 months),7 patients survived at the end of follow-up.The local control rate (LC) was 79.2％,and 1-and 2-year overall survival rate(OS) were 82.7％ and 48.6％,respectively.There was a close corelation between the size of lesion and the LC.The LC (PR + CR) was much better at the size of lesion less than 14 cm3 than that at the size more than 65 cm3(x2 =4.17,P＜0.05).When the size was more than 180 cm3,the LC was zero.Toxicity included fatigue (60.7％),grade 1 and 2 digestive system toxicity (28.6％),a transient grade 1 and 2 bone marrow suppression (46.4％) and a transient increase in transaminase(17.8％).No grade 3 toxicity and above and late toxicity were observed.Conclusions Stereotactic body radiation therapy could be suggested as the first choice for the selected patients who suffer form colorectal liver metastases,especially for those who cannot undergo surgery.

Objective To set up a new diagnostic platform based on microarray exon-capture and next-generation sequencing for detecting small mutations in dystrophin gene.The sensitivity and specificity of the method were assessed in clinical settings and the distribution of small mutations in Chinese Duchenne muscular dystrophy/Becker muscular dystrophy (DMD/BMD) patients were also analyzed.Methods Forty-one DMD/BMD patients diagnosed by the clinical criteria without large deletion or duplication (≥ 1exon) were recruited from Peking Union Medical College Hospital consecutively.Genomic DNA was extracted from blood samples.The libraries were prepared.Then exon and intron-exon flanking sequences of DMD gene were captured by custom microarray.Targeted next-generation sequencing and Sanger Sequencing were conducted.The patients who were not detected any disease-causing mutation were performed muscle biopsy.Results Thirty-eight subjects were detected small mutations in DMD gene.All single nucleotide variants (SNVs) and insertion & deletions (INDELs) were validated by Sanger sequencing.Twenty-one novel mutations were reported.The distribution of SNVs and INDELs was similar to other international DMD databases.Upon immunohistochemistry staining of dystrophin protein,1 of 3 mutation-undetected patients was diagnosed as DMD,2 of them were excluded.The specificity of the method was 100％,while the sensitivity was 97.4％.Conclusions Our microarray-captured next-generation sequencing assay could detect SNVs and INDELs with high sensitivity and specificity.Its advantages are economic,time-saving and stable.The platform is suitable for clinical gene diagnosis.