OBJECTIVE:To establish the quality standard for Qiwei kangti powders.METHODS:Radix Astragali,Angelica sinensis,Salvia miltiorrhiza were identified by TLC.The content of tanshinone Ⅱ A(the chief content in Salvia milti-orrhiza) was determined by HPLC.RESULTS:The TLC spots developed were fairly specific.Tanshinone Ⅱ A showed a good linear relationship at a range of 5.44～32.64 ?g?mL-1(r=0.999 8),and its average recovery was 99.54%(RSD=0.955%,n=9).CONCLUSION:The established quality standard is applicable for the quality control of Qiwei kangti powders.

Objective To establish a flow cytometry-based test for the detection of pyroptosis of murine bone marrow-derived macrophages (BMDM). Methods Bone marrow cells were isolated from wild type (WT) C57BL/ 6 mice and/ or caspase-1-/ - C57BL/ 6 mice and then stimulated with macrophage colony-stimulating factor (M-CSF) to differentiate into murine BMDM. PBS, LPS and LPS+adenosine triphosphate (ATP) were respectively used to stimulate the BMDM. Western blot assay was performed to detect the cleav-age of IL-1β and caspase-1. The levels of IL-1β in the supernatants of cell culture were measured by ELISA. Lactate dehydrogenase (LDH) released in the culture media was detected by using LDH kit. The pyroptosis of murine BMDM was detected by using flow cytometry analysis after double-staining with TMR red+caspase-1, AnnexinⅤ+caspase-1 and propidium iodide (PI)+caspase-1. Results IL-1β was detected in the culture medium of BMDM treated with LPS+ATP and the cleavage of IL-1β and caspase-1 was confirmed by Western blot assay, which indicated that the NLRP3 inflammasome was activated by LPS+ATP treatment. Compared with the caspase-1-/ - mice group, higher levels of LDH were detected in the culture medium of BMDM isolated form the WT mice. Results of the flow cytometry analysis after staining BMDM with caspase-1 plus AnnexinⅤ or PI showed that more cells undergoing pyroptosis were detected in the LPS+ATP treat-ment group than that in LPS or PBS treatment group, which were consistent with the results of the reported flow cytometry with caspase-1+TMR red staining. Conclusion The flow cytometry-based test with double-staining of caspase-1 plus AnnexinⅤ or PI could be used for the detection of pyroptosis of murine BMDM.

ABSTRACT:Objective For preparation of vascularized islets , to isolate and culture human adipose derived stem cells , investigate the role of adipose derived stem cells (ADSCs ) in promoting the proliferation and vascularization of human umbilical vein endothelial cells (HUVECs ) co‐cultured in vitro , and explore its mechanism .Methods ADSCs and HUVECs were isolated by collagenase digestion method ,then cultured ,and identified by morphology ,immunofluorescence or multi‐directional differentiation .The co‐culture system of ADSCs and HUVECs was established , HUVECs cultured alone were set up for control group . The proliferation , vascularization and concentration of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b‐FGF)in the supernatant were compared between the two groups .Results The third generational ADSCs had uniform long spindle fiberous morphology and multi‐directional differentiational function . Immunofluorescence test of surface antigens on ADSCs revealed CD44/CD49d (+ ) ,CD31/CD34 (-) ,on HUVECs CD31/vWF (+ ) . High vascular density was found when co‐cultured in Matrigel of ADSCs and HUVECs than alone of HUVECs .Growth curve shown at days 3 , 4 and 5 of the logarithmic phase , HUVECs count in co‐culture group of ADSCs and HUVECs was (4 .52 ± 0 .31) × 104 ,(7 .18 ± 0 .45) × 104 ,and (8 .23 ± 0 .36) × 104 under indirect co‐culture condition , while that in individual HUVECs group was (2 .71 ± 0 .25) × 104 ,(4 .87 ± 0 .26) × 104 ,and (6 .86 ± 0 .33) × 104 ( P<0 .01) .Population doubling time of HUVECs was shorter in co‐culture group than in individual group .Also ,the OD value of HUVECs was higher in co‐culture group than in individual group when cultured at days 1 ,3 ,5 and 7 ( P<0 .01) .When cultured at days 3 ,7 and 13 ,the concentration of VEGF and b‐FGF in the supernatant was higher in co‐culture group than in individual group ( P< 0 .01 ) . Conclusion ADSCs can promote the proliferation and vascularization of HUVECs in vitro co‐culture conditions by secreting or increasing the HUVECs secretion of VEGF and b‐FGF .

Objective To investigate the application of 3D reconstruction system in the clinical departments of the hospital.Methods 3D reconstruction system had its architecture,main features,working flow and principle of auto preprocessing software introduced,and then applied to auto preprocessing of PACS images to realize auto reconstruction,which had the function of 3D post processing.Results 3D reconstruction system gifted the doctor in clinical departments with the access to the images and after treatment,and changed the traditional working mode in imaging department.Conclusion Fusion imaging has 3D reconstruction as the main technique,which eliminates the deficiency in reading radiological images and innovates medical service mode.

Objective:By exploring the epidemic level and related factors of depressive symptoms,to provide the references for improving mental health of the physical disabled.Methods:The data were from the 2013 China Health and Retirement Longitudinal Study database.Persons with self-reported physical disabled were selected as subjects,and a total of 974 valid ones were obtained,with the average age of (62 ± 10) yeats.The depressive symptoms were measured with the Center for Epidemiologic Studies Depression (CES-D),with the cut-off score of equal or higher than 10 as having depressive symptom.The independent variables included age,gender,education level,marital status,urban and rural areas,self-rated general health,self-rated heating,self-rated memorizing,life satisfaction,smoking,drinking,sleep duration,chronic diseases,activity of daily life,social activity,family support,and disability periods.Chi-square test was used to analyze the effect of each factor on prevalence of depressive symptoms.Binary logistic regression was used to analyze the effect of multi-factors.Results:The prevalence of depressive symptom among the physical disabled was 46.1％.The subjects who were female,single,suffered from chronic disease,without family support,short-time sleeper were more likely to have depressive symptoms than the controls (OR =1.35,1.62,1.60,1.67,2.58;P ＜0.05).The subjects who had better self-rated general health,better self-rated hearing,better self-rated memorizing,better life satisfaction were less likely to have depressive symptoms than the controls (OR =0.38,0.53,0.47,0.09;P ＜ 0.05).Conclusion:It suggests that the prevalence of depressive symptoms is higher among the physical disabled aged 45 years of age or older.It should take appropriate measures to reduce emotinal problems for them.

OBJECTIVE:To establish a method for the determination of dissolution of Xiaocaihu pill,and compare the differ-ence of preparation from different manufacturers. METHODS:Using 0.1 mol/L HCl as dissolution medium,rotating basket method was used to determine the dissolution of preparations. HPLC was adopted to determine the content of baicalin:column was TSKgel ODS C18 with mobile phase of methanol-water-phosphoric acid (65∶35∶0.7,V/V/V) at a flow rate of 1 ml/min,detection wave-length was 280 nm,column temperature was 30 ℃,and injection volume was 5 μl. RESULTS:The linear range of baicalin was 0.488-124.8 mg/L (r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2.0%;recovery was 100.14%-104.78%(RSD=1.58%,n=9). The average t50(50% dissolution time)of baicalin was 85.81 min. CONCLUSIONS:The method is simple with good precision,stability and reproducibility,and can be used for the dissolution determination of Xiaocaihu pill. Xiaocaihu pill from different manufacturers shows great differences,both preparation formulation and clinical use should attach importance to the dissolution of solid preparations.

Objective To investigate the correlation between immune cell function and the infection after renal transplantation through monitoring of immune function intracellular ATP by Cylex ImmuKnow assay,and explore its significance in individual immunosuppressive therapy of renal transplantion recipients.Method We collected 44 renal transplant patients suffered from pulmonary infection from January 2014 to March 2015.The patients were divided into two groups according to the clinical status,namely,ImmuKnow monitoring group (n =22) and empirical treatment group (n =22).Thirty-two non-infection recipients were collected as controls.All the kidney transplantation recipients received immunosuppressive therapy based on calcineurin inhibitors,mycophenolate mofetil and prednisone,and ATG for induction therapy after transplantatior.The immune cell function levels were measured by Cylex ImmuKnow assay.The whole blood samples were collected before infection onset,at the time of infection,and 1 week after infection resolution.Result When infection occurred,ATP concentrations in CD4+ T cells of the kidney transplant recipients were significantly lower than those in non-infection group [(151.30--71.35 ng/mL vs.(308.34 ± 141.29 ng/mL,P＜0.05).When the infection got controlled,the ATP concentrations in CD4+ T cells increased to those before infection occurred.The average hospitalization time in ImmuKnow monitoring group was 12.27 ± 0.74 days,which was significantly shorter than in empirical treatment group (16.64 ± 1.98 days,P＜ 0.05).The incidence of acute rejection was 4.5％ in ImmuKnow monitoring group,and 13.6％ in empirical treatment group (P＞0.05).Conclusion The examination of ATP in CD4+ T cells by Cylex Immuknow assay could reflect the status of cellular immunity,provide reliable and objective basis for the diagnosis and treatment of infection after renal transplantation,and guide the clinical individualized immunosuppressive therapy.

Objective To isolate and culture human adipose-derived stem cells (hADSCs),investigate the influence of hADSCs on the cellular morphology,survival rate,and function of human islet cells under the in vitro non-contact co-culture conditions,and explore its mechanism.Method hADSCs were isolated by collagenase digestion method,then cultured,and identified by morphology,immunofluorescence and multi-directional differentiation.Adult islet cells were separated and purified by Liberase enzyme and Ficoll 400,then divided into co-culture group and individual group.The cellular growth morphology of islet cells was observed by inverted phase contrast microscope.The survival rate of islet cells,insulin secretory volume,insulin stimulation index and concentration of growth factor in the supernatant were compared between the two groups.Result hADSCs of the third generation showed uniform long spindle fibrocyte-like morphology,and had multi-directional differentiational potentials of osteogenesis and adipogenesis.Immunofluorescence test of surface antigens on hADSCs revealed CD44 + and CD49d +,CD31-,CD34-and CD106-.After 14-day culture,the islet cellular morphology in co-culture group was more intact than that in individual group.The survival rate of islet cells in co-culture group was (82.83 + 2.32) ％,and that in individual group was (53.00 + 2.82) ％ (P＜0.01).Insulin secretory volumes were (23.66 + 2.11) and (7.82 +1.09) mU/L respectively in co-culture group and individual group under high glucose concentration,and 13.22 + 0.77 and 6.40 + 0.44 mU/L respectively under low glucose concentration (P＜0.01 for all).Insulin stimulation index was decreased from 1.67 + 0.10 (at 3rd day) to 1.77 + 0.13 (at 14th day) in co-culture group,and from (1.67 + 0.10) (at 3rd day) to (1.77 + 0.13) (at 14th day) in individual group (P＜0.01).After 14-day culture,the concentrations of HGF,TGF-β,VEGF and bFGF in the supernatant were higher in co-culture group than in individual group (P＜0.01).Conclusion hADSCs were isolated and cultured successfully from adult adipose tissue.They could increase the survival rate and improve the function of islet cells when co-culture with the adult islet cells in vitro through secreting HGF,TGF-β,VEGF and b-FGF.

Objective To assess the left atrial early function in pilots with essential hypertension by strian rate imaging (SRI). Methods Twenty-five hypertensive pilots without left ventricular hypertrophy and expansion (LVN) were selected. Twenty-ifve healthy pilots were included as control group. At the left ventricular apical four-chamber view, three-chamber view, two-chamber view, the systolic, early and late diastolic peak strain rate (SRs, SRe, SRa) curves were acquired by SRI in the basal section, the middle section and the upper section of the wall of left atrium. The average strain rate (mSRs, mSRe, mSRa) was calculated. The LA maximal, minimal and pre-systolic volume (LAVmax , LAVmin and LAVp), LA passive ejection fraction (LAPEF), LA active ejection fraction (LAAEF) , left atrial stroke volume (LASV), E/A and E/E′ were calculated. Results The LAPEF, LAAEF, LASV, E/A, E/E′were (38.96±6.43)%, (61.11±5.86)%, (14.74±5.33)mm, 1.23±0.08, 8.63±1.77 in the control group, and were (34.18±5.72)%, (63.23±6.89)%, (10.12±3.23)mm, 1.04±0.26, 7.16±0.84 in the study group. Compared with the control group, signiifcant difference was detect in LAPEF, LAAEF, LASV, E/E′(t=4.840, 2.690, 4.801, 2.950, all P<0.05). No signiifcant difference was found in E/A between the study and control group (t=-1.424, P=0.178). There was signiifcant positive correlation between SRa and E/E′(r=0.632, P<0.05). Conclusion SRI could be applied in the evaluation of early left atrial function. In pilots with essential hypertension, the changes of strain rate in left atrial wall, which is prior to the modeling geometry of left ventricular, could be detected by SRI.

Objective To analyze Holder pasteurization,temperature and duration of frozen,duration of continuous breast milk pumping and types of pumping tube on macronutrients and total energy in breast milk and the circadian variation of macronutrients and energy content of breast milk.Methods Two hundred and two breast milk samples were collected from healthy mothers of 188 infants who were hospitalized in Department of Pediatrics,Obstetrics and Gynecology Hospital Affiliated to Nanjing Medical University from April 2016 to December 2016.Fat,protein,carbohydrate and energy contents of the breast milk samples were detected both before and after various processes to analyze the influence of Holder pasteurization,frozen storage conditions (temperature and time),duration of continuous pumping and types of pumping tube and to observe the variations within a single day.Paired t-test,repeated-measures analysis of variance,one-way analysis of variance and LSD-t test were used for statistical analysis.Results The contents of fat,protein and energy in breast milk were significantly decreased [(3.69 ± 0.98) vs (4.22 ± 1.00) g/dl,t=9.405;(2.12 ± 0.34) vs (2.21 ± 0.34) g/dl,t=4.785;(69.77±9.35) vs (74.94±9.53) kcal/dl (1 kcal=4.184 kJ),t=9.470],while the carbohydrate content was increased [(6.66±0.17) vs (6.62±0.17) g/dl,t=-3.442] after Holder pasteurization comparing with those before (all P＜0.01).Breast milk frozen at =80 ℃ had higher protein content than that frozen at-20 ℃ (Ftemperature=4.990,Ptemperature=0.027).Breast milk stored at-20 ℃ for 7,15,30,60 and 90 days showed significantly decreased fat,protein and energy contents as compared with those in fresh breast milk,while the carbohydrate contents in the breast milk samples stored at-20 ℃ for 15,30 and 60 days were significantly increased and that kept for 90 days were lower than those at 30 and 60 days (LSD-t test,P＜0.05).Compared with fresh breast milk,breast milk stored at-80 ℃ showed significantly decreased fat,protein and energy contents at 60 d and 90 d,but increased carbohydrate content at 60 d (LSD-t test,P＜0.05).Compared with fresh breast milk,breast milk which was pumped at 1,2 or 3 h through gastric tube (size 6,8 or 10) or transfusion connecting pipe had lower fat content (LSD-t test,P＜0.05).Compared with fresh milk,breast milk had lower energy content on condition that it was pumped through gastric tube size 6 at 2 or 3 h or through gastric tube size 8 or 10 or transfusion connecting pipe at 1,2 or 3 h (LSD-t test,P＜0.05).Breast milk secreted from 12:01 to 15:00 had higher fat and energy contents [(5.16±0.93) g/dl and (86.96±9.35) kcal/dl] than that secreted from 00:01 to 03:00[(4.54±0.75) g/dl and (80.77±7.75) kcal/dl],from 03:01 to 06:00 [(4.61 ±0.75) g/dl and (81.44±7.69) kcal/dl] or from 21:01 to 24:00 [(4.71 ± 0.88) g/dl and (82.44± 8.82) kcal/dl].Moreover,breast milk secreted from 09:01 to 12:00 had higher fat and energy contents [(5.01 ± 0.94) g/dl and (85.31 ± 9.62) kcal/dl] than that secreted from 00:01 to 03:00,and that secreted from 09:01 to 12:00 had higher fat contents than that secreted from 03:01 to 06:00 (LSD-t test,P＜0.05).Conclusions Fresh breast milk is better than processed milk to meet the infantile need for growth and development.