In the present study, the regulation of Vitreoscilla hemoglobin (VHb) on astragaloside IV biosynthesis was investigated. An intermediate expression vector consisting of the CaMV35S promoter fused to the vgb and nopaline synthase terminator was transferred into Astragalus membranaceus via Agrobacterium rhizogenes. The transgenic hairy roots were confirmed by PCR amplification and Southern blot hybridization. The expression of vgb in transgenic hairy roots was confirmed by RT-PCR. After 15 days cultivation, the dry weight and growth rate of transgenic hairy roots were higher than that of the non-transgenic hairy root. ELSD-HPLC analysis showed that astragaloside IV content of transgenic hairy roots was 5 to 6 times of non-transgenic hairy root control and 10 to 12 times of Radix Astragali from Shanxi Province. These results suggested that the expression of vgb promoted the growth of transgenic hairy roots, and increased the content of astragaloside IV.

Objective To detect and separate the side population cells(SP) from multiple myeloma (MM) cell lines PRIM8226,and to study their biological characteristics.Methods Fluorescence activated cell sorter (FACS) and Hoechst33342/PI dye were used to sort SP cells of PRIM8226.The multiplication capacity was tested by the growth curve and MTT test,SP cells proportion and the cell cycle were analyzed by flow cytometry,the colony-formtion ability was compared in terms of colony forming experiment,the expression of c-myc,KIF4,SOX2,OCT4 was tested by RT-PCR.The oncogenicity of the cells was analyzed by nude mouse transplantation tumor experiment in vivo.Results The ratio of SP cells in PRIM8226 was (1.78±0.89) ％.More SP cells in the G0 / G1 period,(44.34±3.09) ％ vs (28.49±1.97) ％,P ＜ 0.05,and fewer cells in S phase than MP cells,(38.83±3.69) ％ vs (51.49±4.62) ％,P ＜ 0.05.There were no difference in the expression of CD38 and CD138 between SP cells and MP cells,respectively,(78.5±8.5) ％ vs (82.0±4.0) ％ and (72.3±15.7) ％ vs (84.3±11.9) ％,P ＞ 0.05.Colony formation assay showed that the colony forming efficiency of the SP cells was higher than the MP cells,single cell clone diameter,the number of clone forming,the clone formtion rate were significantly higher than MP cells,0.280±0.016 vs 0.118±0.019,1 722±127 vs 358±14,(86.1±3.46) ％ vs (17.9±1.88) ％,P ＜ 0.05.The mRNA expression levels of c-myc,KIF4,SOX2,OCT4 in SP cells were higher than those in MP cells,c-myc (29.90±3.73) ％ vs (16.84±2.35) ％,KIF4 (29.97±2.89) ％ vs (19.06±1.23) ％,SOX2 (40.00±4.58) ％ vs (16.62±2.09) ％,OCT4 (32.96±0.92) ％ vs (23.27±0.92) ％,all P ＜ 0.05.Nude mouse transplantation tumor experiment in vivo showed the oncogenicity of the SP cells was more stronger than MP cells (5×103 vs 5×l05).Conclusion There are notably difference in the cell cycle,colony formation assay,the mRNA expression levels and oncogenicity,but no difference in the expression of CD38 and CD138,the proliferation ability between SP cells and MP cells.

Objective:To examine whether dysfunctional attitudes and cognitive bias change with fluctuations in depressive symptoms. Methods:122 patients with MDD (major depressive disorder) were assessed with DAS (dysfunctional attitude scale),CBQ (cognitive bias questionnaire) ,and HRSD (Hamilton rating scale for depression) before and after the 8-week treatment. 51 normal subjects were assessed with the DAS, CBQ and SDS as controls.Results:(1) Compared with normal controls, at the baseline MDD patients had significantly higher total DAS scores (158.2?34.6 vs. 119.3?23.4,t=8.559, P=0.000) and the score of CBQ (5.19?3.77 vs. 1.69?1.83, t=8.208, P=0.000). (2) After 8 weeks, the HRSD scores of patients reduced to Median 7.0, and the DAS and CBQ scores significantly lower than those in baseline (DAS total score from 158.2?34.6 to 143.3?29.3, t=3.611, P=0.000; the median of CBQ decreased from 5.0 to 3.0;z=4.670, P=0.000). (3) The DAS score (131.1?25.7) and CBQ score (median 3.0) of full recovery patients with full recovery remained higher than those of controls (t=2.397, P=0.018 ;z=3.990, P=0.000).Conclusion:Dysfunctional attitudes and cognitive bias were influenced by fluctuations in stata or demonstrateing stability over time. They were all state-trait features.

Objective:To determine berberine and palmatine in Phellodendron chinense Schneid.and its granules.Methods: The reversed-phase ion pair high performance liquid chromatography(RP-HPLC) was used and the validation of the method was tested.The chromatography condition was with Lichrospher C18 column(4.6 mm?250 mm,5 ?m), mobile phase was ACN∶25 mmol/L NaH 2PO 4∶25 mmol/L SDS(2∶1∶1),flow speed was 1.0 ml/min,detection wavelength was 345 nm,and temperature of column was 25℃,Phellodendron chinense Schneid.and its granules were extracted with methanol solution of hydrochloric acid.Results: The theoretical plate number of berberine and palmatine were 14 906 and 14 847,the resolution were 2.33 and 2.86,the tailing factor were 1.09 and 1.06,respectively; all the parameters were suitable for determination.The calibration curves were linear in the range of 40-500 ng,Y=698 278X-3 846,r=1.000(berberine) and 20-250 ng, Y= 536 632X- 7 738, r=0.999 9,r=0.999 4(palmatine).The intra-day and inter-day precision(RSD) at low,middle and high injection amount was all less than 2.5%(berberine) and 1.5%(palmatine).The stability(RSD) was 0.66%(berberine) and 0.70% (palmatine) in 48 h.The recurrence(RSD,n=5) was 0.11%(berberine) and 0.12%(palmatine).The limits of detection was 2.0 ng(berberine) and 1.0 ng(palmatine).The recoveries were 100.4% (RSD=0.12%,n=3) for berberine and 99.80%(RSD=0.22%,n=3) for palmatine.The contents of berberine and palmatine in 3 batch of Phellodendron chinense Schneid.and 5 batch of its granules were determined.Conclusion: Our method can be used for determination of berberine and palmatine in Phellodendron chinense Schneid.and its granules, which is simple and reliable.

Objective:To analyze influencing factors for sarcopenia in people of advanced age, in order to provide insight and evidence for the prevention and treatment of sarcopenia in people belonging to this age group.Methods:Data from 167 people of advanced age seeking care at our department from December 2014 to July 2017 were retrospectively analyzed.According to the diagnostic criteria for sarcopenia of the Asian Working Group for Sarcopenia, subjects were divided into the sarcopenia group( n=46, 27.5%)and the non-sarcopenia group( n=121). Differences in body composition, energy intake, quantities and proportions of three major nutrients were analyzed between males and females.Related influencing factors for sarcopenia were analyzed by using multiple linear regression. Results:Compared with the non-sarcopenia group, the sarcopenia group had lower body mass index, waist-hip ratio, fat mass, total energy intake and protein( P<0.05)but higher age and fat intake( P<0.05). Values for grip strength, muscle mass, index of skeletal muscle, adjusted muscle mass by body mass index, total energy intake, carbohydrates, fat and protein were lower and the percentage of body fat was higher in females than in males( P<0.05). Multiple Logistic regression analysis showed that insufficient intake of protein( β=-0.290, OR=0.748, 95% CI: 0.569-0.984, P<0.05), reduction of body fat mass( β=-2.673, OR=0.069, 95% CI: 0.010-0.488, P<0.05)and excessive visceral fat accumulation( β=0.739, OR=2.094, 95% CI: 1.219-3.597, P<0.01)were correlated with sarcopenia in people of advanced age. Conclusions:The occurrence of sarcopenia is higher in people of advanced age and is related to insufficient intake of protein, reduction of body fat mass and excessive visceral fat accumulation.Individualized nutrition evaluation and support should be carried out as early as possible for people in this age group.

Objective:To investigate the muscular quality and its related influencing factors in elderly with sarcopenic obesity.Methods:The internalized 696 elderly subjects meeting inclusion and exclusion criteria were divided into four groups including the sarcopenic obesity(n=55), sarcopenia(n=8), simple obesity(n=481)and normal control(n=152)groups.The intergroup difference was retrospectively analyzed in the parameters of body composition, dietary intake of total energy and three major nutrients and their proportions.Multiple linear regression analysis was used to analyze the influencing factors related to the occurrence of sarcopenic obesity in the elderly.Results:The sarcopenic obesity group versus normal control group showed a higher level or value in age, body mass index, waist-hip ratio, fat mass, percentage of body fat, visceral fat area, fasting blood glucose, estimated glomerular filtration rate( P<0.01 or 0.05), and showed a lower level or value in grip strength, skeletal muscle, skeletal muscle index, muscle quality(grip strength/limb skeletal muscle), intake of energy, carbohydrate, fat and protein, body weight-adjusted intake of energy and protein, as well as ideal body weight-adjusted intake of energy and protein( P<0.01). The detection rate of sarcopenic obesity was 7.90%(n=55), including 7.60% in males(n=48)and 11.48% in females(n=7 cases). The detection rate of sarcopenic obesity was increased along with increasing age with 0.07%(2/287)in 60-69 years old group, 4.94%(12/243)in 70-79 years old group, and 24.70%(41/166)in ≥80 years old group( χ2=87.76, P<0.01). Taking the median point of grip strength/limb skeletal muscle volume as the cutoff point, the decrease rate of muscle quality was 12.36%(86/696)in total elderly subjects, 2.63%(4/152)in the control group, 11.64%(86/635)in the obesity group, 37.5%(3/8)in the sarcopenia group and 41.82%(23/55)in the sarcopenic obesity group, with an increasing trend of the decrease of muscle quality, which had significant differences( χ2=62.25, P<0.01). Multiple logistic regression analysis showed that aging, excessive visceral fat area, insufficient protein intake and decreased basal metabolism were the independent risk factors for sarcopenic obesity in elderly people( P<0.01 or P<0.05). Conclusions:The incidence of decrease of muscle quality is higher in the elderly with sarcopenic obesity than other elderly groups.Sarcopenic obesity is correlated with aging, insufficient intake of protein, decreased basal metabolism and excessive visceral fat accumulation.Individual evaluation and support is necessary in elderly people with sarcopenic obesity.

Objective To compare the effects of three kinds of fixative solutions on paraffin section of mouse lens tissue and optimize the fixing?method of paraffin section in mouse lens tissue. Methods Three kinds of conventional fixatives were selected for the test ,including the conven?tional Davison’s solution,modified Davison’s solution and 10%neutral buffered formalin. Mice eyeball tissues were fixed with three different fixa?tives,embedded,sliced and then stained with HE method. The paraffin slices were observed under the light microscope. Results The structures of lens and retina fixed in conventional Davison ’s fixative solutions were clear and intact ,and the cells were arranged regularly and compactly. There was no eyeball distortion,contraction and retinal detachment in the eyeballs fixed in modified and conventional Davison’s fixative solution. However,the ones fixed in 10%neutral buffered formalin showed eyeball distortion and contraction,space and spherules. Conclusion The mice lens slides made from tissues fixed by conventional Davison ’s fixative solution are better than fixed by modified Davison ’s fixative solution and the 10%neutral buffered formalin fixed ones.

Objective: To observe the effect of telomerase-antisense DNA on apoptosis of hepatoma cells induced by arsenic trioxide, in an effort to look for a new anti-hepatic cancer agent with high efficiency, low cytotoxicity. Methods: We designed and synthesized a 20nt telomerase-antisense DNA targeting telomerase template and observed its influence on the telomerase activity of hepatoma cells. H-E staining, flow cytometry, and DNA agarose electrophoresis were used to study the preventive effect of telomerase-antisense DNA on hepatoma cells apoptosis induced by arsenic trioxide. Flow cytometry was used to detect the expression of Fas, Fas-L, and bcl-2. Results: Telomerase-antisense DNA (5 ?mol/L) effectively inhibited the telomerase activity of hepatoma cells after 24 hours (P

Objective To investigate reliability and validity of the Chinese version of the schedule for assessment of insight (SAI).Methods The original version of SAI was translated into Chinese and then backtranslated into English to develop the Chinese version of SAI which reliability and validity were tested.220 subjects who met the diagnosis of Schizophrenia,schizophreniform disorder,schizoaffective disorder,delusional disorder or brief psychotic disorder were assessed.Results The Cronbach's α was 0.890 ; the test-retest reliability was 0.742 ; the inter-rater reliability was 0.986.Three main factors were obtained from the factor load matrix as awareness of illness,ability to relabel psychotic experience,compliance with treatment.The loading coefficient were 0.883-0.944.The correlation of the SAI total score and three subscales with insight of PANSS were-0.635,-0.305,-0.631 and-0.512 respectively (P ＜ 0.01).Conclusion The SAI has good reliability and validity.The scale is useful to measure insight in psychosis.