Objective To confirm the best dry technics,the colormetry of anthrone-sulfuric method was employed to determine the content of polysaccarides in fresh water mussel and three factors of temperature,dry time and tiled destiny were compared.Method Colorimetry of anthrone-sulfuric acid.Results and ConclusionThe best dry technics was: 2.5 kg/m2 tiled destiny,temperature at 90 ℃ and drying within 2 hours.The content of total amylase in polysaccharides in fresh water mussel was 21%.The method appeared to be reliable and operable for quality control.

Objective To investigate the change of peripheral blood CD34+ cells and white blood cells(WBC)count in the patients with brain traumatic injury in convalescence stage during hyperbaric oxygen(HBO)treatment.Methods The peripheral blood CD34+ cells and WBC count were measured in 11 patients with brain traumatic injury in convalescence stage who accepted HBO treatment before and 1,2,3,4,and 5 weeks during the treatment.Other 11 persons were measured once as controls.Results Compared with the controls,the number of CD34+ cells showed normal in the 1st and the 3rd week,but increased in the 2nd and the 5th week in the HBO group(P<0.05).It was increased in the 5th week compared with that in the 1st and the 3rd week(P<0.05)in the treatment group.The number of WBC showed no significance and the patients' symptoms improved during the HBO treatment.Conclusion HBO can mobilize bone marrow derived stem cells marked by CD34,which may participate in the restoration of brain traumatic injury in convalescence stage.

BACKGROUND: Reports regarding adipose-derived stern cells (ADSCs) differentiation into dopaminergic (DN) neurons are few in addition, there is not experimental evidence of the effect of ADSCs on maintaining the survival of DN neurons.OBJECTIVE: To investigate the effect of glial cell-derived neurokophic factor (GDNF)-modified adipose-derived stem cells on survival of DN neurons under co-cultured condition.DESIGN, TIME AND SETrlNG: The in vitro cytology experiment was conducted at the Institute of Otolaryngology-Head and Neck Surgery and Key Laboratory of Zoonoses of Ministry of Education between March and December 2007.MATERIALS: Wistar rats with 3-weeks-old, or 14 days of pregnancy were provided by Norman Bethune College of Medicine, Jilin University.METHODS: The GDNF recombinant adenovirus was constructed by using pAdTrackCMV and pAdEasy-1 system. DN neurons were obtained from the rostral mesencaphalic tegmentum of Wistar rat embryos by using trypsin and collagenase method. ADSCs isolated from rat inguinal fat pads were digested with collagenase Ⅱ, cultured and passaged in vitro. When the cells reached 60% cenfluency at the 3rd passage, cells were transfectad with 1×109vp/mL of Ad-GDNF for 1 hour and then transferred into growth medium for another 24 hours, and GDNF level in cell supematant was detected by ELISA assay. Meanwhile, the co-cultured of ADSCs and DN neurons were carried out for following 7 days. With GFP-modified ADSCs was served as a control group.MAIN OUTCOME MEASURES: The effect of co-cultured condition on the survival of DN neurons, as well as the differentiation of GDNF-modified ADSCs was detected by immunofluorescence staining.RESULTS: GDNF appeared in ADSCs supematant at 24 hours after Ad-GDNF transfection and reached a peak at 72 hours.There was approximately 80% GFP-positive labeled in ADSCs. The tyrosinase hydroxylase staining results demonstrated that the rate of survival DN neurons were significantly increased than in DA neurons cultured alone, co-cultured group of GFP-modified ADSCs and GDNF-modified ADSCs groups (55%, 15%, 25%, P < 0.01). However, there were no co-expressing TH and GFP positive cells appeared at 7 days of co-culture, which indicated that the co-cultured condition was not available to ADSCs differentiation.CONCLUSION: The co-cultured of GDNF modified ADSCs and DN neurons can promote the survival and growth of cultured DN neurons, however, it can not induce ADSCs differentiate into DN neurons.

Objective To evaluate the efficacy of surgical excision combined with iodine-125 implants intmcavitary radiotherapy for neumglioma Method After excision,27 patients with nenroglioma were implanted with the iodine-125 into the tumor bed or the residual tumor tissues.Results Of all cases,there were no operative mortality and no serious complication.The survival time of 25 cases exceeded 6 months and 1 glioblastoma recurrence,22 cases exceeded 12 months,3 glioblastoma and 1 astrocytoma Ⅲgrade recurrence.There were 19 cases exceeding 24 months,15 cases no recurrence,3 cases died for cere bral hernia because of giving.up further tremment,1 case surviving.Conclusions Surgical excision Combined with iodine-125 implants intracavitary radiotherapy can improve partial control rate,prolonged the survival time of patients with neuroglioma,and complications is rare.It is an efficient therapy for neu roglioma

Objective To study the effects of fat grafting on partial jaw deformity.Methods A total of 19 cases of partial jaw deformity.were used fat grafting in past 2.5 years,from January 2012 to June 2015.All analyses were to find suitable fat grafting method for partial jaw deformiw.Fat graft was obtained by low pressure suction and low speed centrifugation.Results A lot of 15 patients in this group had been followed up for 6 months to 2 years.9 patients was satisfied with once fat grafting and 6 with twice.Conclusions Fat grafting provides a useful alternative to partial jaw deformity with normal occlusion.But it also remains to be seen what the forward effects of fat grafting on partial jaw deformity are.

OBJECTIVE:To investigate the long-term toxicity of repeated intramuscular injection of Escherichia coli O157∶H7 polysaccharide-conjugated vaccine(O157) in rats so as to provide safety evidence for clinical trials.METHODS:A total of 48 SD rats were randomly assigned to receive either 0.5 mL vaccine(containing 25 ?g polysaccharides) (immunization group,n=24) or phosphate buffered solution (PBS,control group,n=24) with the same volume for 3 times at a dose interval of two weeks.Sacrifice of 6 rats in each group were scheduled at 2 weeks after first immunization,and at 1,3,and 5 weeks after the third immunization,respectively for observation and determination of hematological and biochemical parameters,histopathology,specific antibody,myeloid tissue,the tissues in injection sites,etc.RESULTS:Compared with control group,immunization group showed no significant pathological change except the dynamic regular change of some hematological parameters induced by the immunization,and no immunologic system damage was observed.CONCLUSION:Repeated intramuscular injection with O157 vaccine in rats wouldn't cause overt toxicity and local irritation.

BACKGROUND: The abnormality of the function of arcuate nucleus may be an important factor of obesity. It has been known that the mechanisms of acupuncture in treating obesity are related to nervous and neurohumoral regulation. What is the regulating effect of acupuncture on the function of arcuate nucleus?OBJECTIVE: To study the effects of acupuncture on the function of arcuate nucleus of obese rats, and further investigate central nervous functional mechanism of reducing weight by acupuncture.DESIGN: Randomized controlled study based on the experimental animals.SETTING: Acupuncture institute in second clinical medical college of a university of traditional Chinese medicine, and a population management college.MATERIALS: This experiment was carried out in the Acupuncture Institute of Second Clinical Medical College, Nanjing University of Traditional Chinese Medicine between April and October 2002. One-month old male SD rats just in ablactation were selected.METHODS: Rats fed with ordinary wholesome rat-feed were in the normal group. The successfully established experimental obese rats models were randomly divided as control group and acupuncture group with 12 rats in each group. Rats in the acupuncture group were given acupuncture treatment for 14 days, and rats in the normal and control groups were put into rat fixation-machine for 15 minutes every day, lasting for 14 days. Body mass, Lee' s index, body lipid, level of central and peripheral leptin and insulin(INS) as well as the frequency of spontaneous discharge of nerve cell in the arcuate nucleus(ARC) of hypothalamus in obese rats were observed with nervous electrophysiological and nervous biochemical technology before and after acupuncture.MAIN OUTCOME MEASURES: ① Effect of acupuncture on obesity index, fat contents of pericardium, kidney and epididymis of experimental obese rats. ② Effect of acupuncture on the frequency of spontaneous discharge of ARC nerve cell of hypothalamus in obese rats. ③ Effect of acupuncture on the contents of hypothalamic and serum leptin and INS of rats in each groups. ④ Correlation of the frequency of spontaneous discharge of ARC nerve cell of hypothalamus with the level of hypothalamic leptin and INS.RESULTS: The body mass[(517.74±29.35) g], Lee' s index (319.85 ± 3.96) and body fat content of obesity rats [ (13.88 ± 1.32) g] were all significantly higher than those in the normal rats, the differences were significant(P ＜ 0.01); the frequency of spontaneous discharge of nerve cell in ARC of hypothalamus in obese rats [ (3.12 ± 1.92) Hz] were all markedly lower than those of the normal rats[ (8.99 ± 2.71 ) Hz], the differences were significant (P ＜ 0.01 ); the frequency of spontaneous discharge of nerve cell in ARC was negatively correlated with body mass, Lee' s index and body lipid( r = -0. 592, -0. 672, -0. 521 ) . Acupuncture significantly increased the frequency of spontaneous discharge of nerve cell in ARC of hypothalamus in obese rats [(9.75 ±2.02) Hz] as compared with the control group, the differences were significant( P ＜ 0. 01 ).CONCLUSION: The good regulating effect of acupuncture on the function of arcuate nucleus of hypothalamus might be one of mechanisms in the antiobesity of acupuncture.

Objective:To investigate if acupuncture has an inhibitory effect on fatty toxicity and its possible mechanism. Methods: Thirty-four patients with simple obesity were clinically treated with acupuncture for 3 courses. Before and after treatment, body mass index(BMI) and fat percentage(F%) were determined, and serum insulin and adiponectin were measured by enzyme linked immunoassay, blood lipid and sugar by biochemical colorimetry and eight kinds of free fatty acids(FFAs) by high performance liquid chromatography. Twenty normal persons were selected as a control. Results:The total efficacy rate was 88.2%. Acupuncture could increase the decreased insulin sensitivity index(ISI) in the patients(t=-5.103, P=0.000). The correlation of a decrease in F% with an increase in unsaturated fatty acid after acupuncture was of significance(r=0.402, P=0.019) and its correlation with an increase in the ratio between unsaturated fatty acid and saturated fatty acid was also of significance(r=0.348, P=0.044). The correlation between increases in high density lipoprotein and in eicosapentaenoic acid was of significance(r=0.352, P＝0.041). The correlation between a decrease in BMI and an increase in high density lipoprotein was of significance(r=0.357, P=0.038). Adiponectin level significantly rose after acupuncture (compared with pre-treatment, P=0.000). The correlation between a decrease in FFAs and an increase in adiponectin was of significance(r=-0.349, P=0.043). Conclusion: Acupuncture can lower the levels of free fatty acids in the patients and increase their sensitivity to insulin to inhibit the fatty toxicity. The inhibitory effect of acupuncture on fatty toxicity is somewhat related to a rise in adiponectin level.

AIM: To investigate the association of osteoprotegerin ( OPG) gene single nucleotide polymor-phisms (SNPs), 163A/G (rs3102735) and 245T/G (rs3134069), with susceptibility to rheumatoid arthritis (RA) in Chinese Han population .METHODS:A total of 205 patients with RA and 171 healthy control subjects were enrolled into this study.Genotyping was performed by polymerase chain reaction-based restriction fragment length polymorphism and subsequently confirmed by DNA sequencing .Odds ratio ( OR) and 95%confidence intervals ( CI) were calculated for the risk genotypes and alleles .RESULTS: OPG gene polymorphisms 163A/G and 245T/G were conformed to the Hardy-Weinberg equilibrium .The statistical differences in the genotypes of AA , AG and GG at 163A/G locus were found in RA and controls.The G allele was associated with an increased risk of RA , with OR of 1.219 (95%CI:1.066~2.339).No significant difference was observed between RA group and control group with respect to genotypic and allelic frequencies of OPG gene 245T/G (P>0.05).CONCLUSION:The OPG gene 163A/G SNP may be associated with RA susceptibility , and G allele may be the risk factor for developing RA .

Objective:To explore the effect of ionizing radiation on apoptosis of lung cancer H460 cells after ATRX was silenced by RNAi and its mechanism.Methods:The lentivirus expression vectors targeting ATRX were transfected into the 293T cells,and the lung cancer H460 cells were infected with lentivirus twice,and the ATRX silenced cell model was obtained after puromycin positive screening,then they were named as sh-ATRX1-H460,sh-ATRX2-H460,and sh-ATRX3-H460 cells;the sh-control-H460 cells were regarded as control cells.The cells were divided into sh-control-H460 group and sh-ATRX3-H460 group,accroding to the silencing results and were irradiated by 0,2 and 8 Gy X-rays.The expression levels of ATRX,poly(ADP-ribose) polymerase 1(PARP1),and caspase-3 proteins were measured by Western blotting method;the apoptotic rate was measured by flow cytometry and AnnexinⅤ-FITC/PI kits.Results:The lung cancer cell model of sh-ATRX3-H460 silenced by ATRX was obtained successfully.After 2 and 8 Gy X-ray irradiation,compared with before irradiation,the expression level of ATRX protein in sh-control-H460 group was increased,while there was no expression of ATRX protein in sh-control-H460 group;compared with before irradiation,the apoptotic rates of cells in two groups were increased(P<0.05 or P<0.01);the apoptotic rate in sh-ATRX3-H460 group was significantly higher than that in sh-control-H460 group after 8 Gy X-ray irradiation (P<0.01).The expressions of cleaved PARP1 in the cells in both two groups after 2 Gy and 8 Gy X-ray irradiation were increased and showed similar rule.The expression level of procaspase-3 protein in sh-control-H460 group had little change,and it was increased significantly in sh-ATRX3-H460 group after 8 Gy X-ray irradiation.Conclusion:ATRX silencing can be achived by RNAi,then the silencing could increase the apoptosis induced by irradiation and its mechanism may be related to the PARP1-caspase-3 pathway.