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 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 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.

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.

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 explore the relationship between adjustment of immunosuppressant and prognosis in renal transplantation recipients with pulmonary infection. Methods The clinical data of 98 patients with pulmonary infection following renal transplantation were retrospectively analyzed.Patients were divided into two groups: conventional group (n = 45) and immunosuppressant adjustment group (n = 53). The mortality, recovery time and rejection rate in two groups were analyzed under the statement of serious infection (SOFA≥12) and slight infection (SOFA＜ 12) by sequential organ failure assessment (SOFA) score. Results When the SOFA scores ≥ 12, the mortality and recovery time in immunosuppressant adjustment group were significantly lower than in conventional group (P＜0.05), but there was no significant difference in the rejection rate between two groups (P＞0.05). When the SOFA scores ＜11, there was no significant difference in mortality and recovery time between the two groups (P＞0.05). The incidence of rejection in immunosuppressant adjustment group was significantly higher than in conventional group (P＜0.05).Conclusion Mortality could be decreased and course of anti-infection treatment could also be shortened by adjusting the immunosuppressant in renal transplantation recipients with serious pulmonary infection (SOFA≥12). Immunosuppressant agent was proposed to maintain the original treatment protocol when the infection was slight (SOFA＜12).

Objective To investigate the influence of proline-rich tyrosine kinase 2 (Pyk2)gene RNA interference on proliferation,invasion and migration of Hep3B hepatocellular carcinoma cells.Methods The Pyk2 gene RNA interference vector was transfected in Hep3B hepatocellular carcinoma cells by lipofectamine. The Hep3B cells divided into three groups:siRNA group (the vector with Pyk2 RNAi gene was transfected), negative control group (the vector without Pyk2 RNAi gene was transfected),and blank control group (no vectors was transfected).Pyk2 mRNA and protein were detected using reverse transcription reverse transcription-poly-merase chain reaction (RT-PCR)and Western blotting.The biological behavior including cell proliferation,inva-sion and migration were detected by 3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide (MTT), transwell and wound healing assay,respectively.Results The expression of Pyk2 mRNA of Hep3B cell line in siRNA group (0.1 6 ±0.03)was significantly decreased than those in negative group (0.74 ±0.1 3)and blank control group (0.77 ±0.1 6),with statistically significant differences (t=51 .46,P=0.000;t=53.21 ,P=0.000).The expression of Pyk2 protein of Hep3B cell line in siRNA group (0.24 ±0.06)was significantly decreased than those in negative group (0.83 ±0.05)and blank control group (0.91 ±0.06),with statisti-cally significant differences (t=57.29,P=0.000;t=68.53,P=0.000).The cell proliferation inhibition rate at 48 hours in siRNA group (26.1 7%±0.28%)was significantly raised than those in negative group (9.28%± 0.22%)and blank control group (6.47%±0.31%),with statistically significant differences (t=31 .45,P=0.004;t=34.64,P=0.002).The number of transmembrane cells in siRNA group (32.5 ±8.5)/1 0 HP was significantly declined than those in negative group (98.4 ±1 2.3 )/1 0 HP and blank control group (1 1 2.6 ± 1 1 .3)/1 0 HP,with statistically significant differences (t=95.64,P=0.000;t=1 05.1 7,P=0.000).The wound healing assay in siRNA group (28.1 7%±1 .46%)was significantly lower than those in negative group (77.38%±2.24%)and blank control group (79.41%±3.1 7%),with statistically significant (t=85.86,P=0.000;t=89.37,P=0.000).Conclusion Pyk2 gene involves the proliferation,invasion and migration of Hep3B cells,which has close correction with development and metastasis of hepatocellular carcinoma.Pyk2 gene is very helpful to become a molecular target for the diagnosis and treatment of hepatocellular carcinoma.