1.Culture and functional identification of macrophages from human peripheral blood
Weiming LIN ; Ailing DAI ; Huifang YIN ; Xiaoyan YANG
Chinese Journal of Immunology 2015;(1):86-89
Objective:To isolate monocytes from human peripheral blood mononuclear cells( PBMC) ,induce macrophages,and identify the function of macrophages.Methods:Monocytes were isolated from PBMC using magnetic activated cell sorting( MACS) anti-CD14 microbead.Sorted CD14+and CD14-cells were checked by flow cytometer to evaluate the efficiency of sorting.The sorted CD14+cells were cultured in IMDM media with 10%human AB serum and 10 ng/ml M-CSF for 7 days to generate macrophages,which were identified by morphological features and phagocytosis function.Results:A high purity of monocytes was obtained by MACS anti-CD14 microbead.The percentage of CD14+cells was 10% and 85.8% before and after sorting, respectively.The macrophages were approximately 40-45 μm in maximum diameter and had the fried egg colony morphological features after 7 days culture.The lymphoma ( Raji) cells were efficiently engulfed by macrophages.Conclusion: The high purity of CD14+monocytes is isolated from PBMC and monocyte-derived macrophages efficiently engulfed lymphoma cells.
2.The anti-cancer effects of physiological deep-sea water combined with hyperthermia for hepatocellular carcinoma in vitro
Youguo DAI ; Weiming LI ; Huirong TANG ; Qian YAO ; Jin CUI
Chongqing Medicine 2016;45(7):899-902
Objective To explore the anti‐cancer effects of physiological deep‐sea water(PDSW) combined with hyperther‐mia for hepatocellular carcinoma in vitro .Methods Deep‐sea water (DSW) from the south Chinese sea was processed ,and made in‐to PDSW ,detection of some elements .In vitro ,the cultured normal liver cells and human hepatoma QGY‐7703 cells were randomly divided into PDSW group and normal saline(NS) group ,the NS group received saline ,the PDSW group received different concentra‐tions of PDSW .Two groups were heated respectively to 6 h of 40 ℃ or 1 h of 43 ℃ ,24 ,48 ,72 h after the administration of PDSW or saline ,the normal liver cells and QGY‐7703 cells proliferation capacity and toxicity were investigated by MTT assay .At the same time testing PDSW and NS in 40 ℃ 6 h for 10 d state of human liver QGY‐7703 cell clone formation rate .Results The results of MTT assay showed that tumor inhibitory rate were time and concentration dependent in tow groups .Tumor inhibitory rate of PD‐SW group in different time was significantly higher than NS group (P<0 .05) .On the other hand ,the inhibitory of hepatocyte for PDSW group in different time were significantly lower than NS group .In addition ,the clone formation rate of PDSW group was lower than those of NS group(P<0 .05) .Conclusion PDSW can improve the heat tolerance of normal liver cells .When combine with heat ,it can obviously inhibit the growth of human liver cancer QGY‐7703 cells .
3.The influence of deep sea water on wound healing of mice
Weiming LI ; Jin CUI ; Pengyuan XU ; Youguo DAI
Chongqing Medicine 2014;(4):462-464
Objective This experiment aimed to study the influence of deep sea water (DSW) on wound healing of mice .Methods 24 Mice were randomly divided into two groups :group DSW(n=12) and group sterilize tap water(STW)(n=12) ,freely feeding for 14 days respectively ,and calculated the amount of food and water .On the 15th day ,1 cm × 1 cm size of wound was established on the back area of mice ,and continued to feed with DSW and STW respectively .Tracking the wound healing rate .Specimen was taken in the edge of wound tissue on postoperative 3 ,5 ,7 days ,then observed histopathological changes .Results Compared group DSW with group STW ,there was no significant difference in the total amount of food and water .5 days after the formation of wounds ,the wound healing rate of group DSW was significantly higher than group STW .Histological observation :compared with group STW ,vascular endothelial cells and new capillaries of the group DSW was increased ,and group DSW had less inflammatory cell and more fibroblast cells proliferation .Conclusion deep sea water can promote wound healing .
4.Effects of omapatrilat on renal tubulointerstitial fibrosis in 5/6 subtotal nephrectomized rats
Shengchuan DAI ; Weiming WANG ; Bin ZHANG ; Nan CHEN
Chinese Journal of Nephrology 1994;0(04):-
Objective To investigate the effect of vasopeptidase inhibitors (VPi) omapatrilat on renal tubulointerstitial fibrosis of 5/6 subtotal nephrectomized rats and to explore new prevention and treatment of renal lubulointerstitial fibrosis.Methods The renal subtotal ablation model was established by surgical 5/6 renal resection in male SD rats.Five animal groups included(1)STN group (subtotal nephreclomy); (2)Omap 10 mg group (10 mg?kg-1?d-1); (3)Omap 40 mg group (40 mg?kg-1?d-1) ; (4)Sham group and (5)Nornial control. Systolic arterial BP and renal function were measured periodically.Animals were sacrificed at the end of the 12th week after operation. Tubulointerstitial injury was scored semiquantitatively by morphological analysis (HE, Masson, PAS and PASM stain). Results Significant tubulointerstitial injury was noted in the STN group compared with normal group and sham group (P
5.Effects of treatment with hydroxythy starch on the serum albumin and immunologic function in postoperative patients with obstructive jaundice
Anyin DAI ; Pengyuan XU ; Dali SUN ; Yunyun CEN ; Xiongzhi CHEN ; Jian LIU ; Weiming LI ; Yuxing QI
Parenteral & Enteral Nutrition 2009;16(4):215-218
Objective: To investigate the effects of trentment with hydroxythy starch(130/0.4) on the serum albumin and immunologic function in postoperative patients with obstructive jaundice.Methods: 24 patients with obstructive jaundice were randomly divided into the control group(n=12) and hydroxythy starch(130/0.4) (HS) group(n=12). The serum ALB was detected 1d, 3d, 5d, 7d after the operation,and the immunologic index were detected 1d and 7d after the operation.Results: The ALB content of control group and HS group were not significantly different in the postoperative 1d and 7 d(P>0.05) and had significant differences in the postoperative 3 d, 5 d (P<0.05).All immunologic index had no significant differences in the postoperative 1d, 7d (P>0.05).Conclusion: The ALB content of patients with obstructive jaundice may decrease postoperatively. Treatment with hydroxylthy starch(130/0.4) can alleviate the drop of the ALB content. But it has no effects on immunologic function.
6.Experimental study on the dynamic changes of cardiomyocyte apoptosis and Caspase-12 activation and on the relationship between the two changes after coronary microembolization in rats
Hanhua ZHU ; Lang LI ; Yi WANG ; Weiming WEN ; Yongguang LU ; Xianming ZHAO ; Rixin DAI ; Tangwei LIU
Chinese Journal of Geriatrics 2010;29(9):755-760
Objective To investigate the dynamic changes of cardiomyocyte apoptosis and Caspase-12 activation after coronary microembolization (CME) in rats. Methods The CME models were produced by injection of 42 μm microspheres (3000/0.1 ml) into the left ventricle during clampinduced ascending aorta occlusion for 10 seconds in adult male Sprague-Dawley rats (CME group).The sham-operation group was injected with saline instead (S group). The survivors were randomly divided into five groups: 3 h, 6 h, 12 h, 24 h and 4 weeks (n=10, each), respectively. In addition,10 rats were designed as normal control group. Cardiomyocyte apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. The expressions of Caspase-3, 12 and procaspase-3 and 12 were detected with Western-blot analysis. The activity of Caspase-12 was determined with fluorometric assay kit. Results (1)Compared with the shamoperation group and normal control group, the apoptosis rates of cardiomyocytes in CME group were significantly increased at each time point respectively (all P<0.05). Apoptotic cardiomyocytes were found mainly in the border zones and infarct foci. The apoptosis rates of cardiomyocytes at 3 h, 6 h,12 h, 24 h and 4 weeks after CME were (1.76±0.68)%, (3.17±1.26)%, (1.34±0.12)%,(1.07±0.65)% and (0.30±0.13)%, respectively. The apoptosis rates of cardiomyocytes increased at 3 h after CME, peaked at 6 h after CME (all P<0.05), and then gradually decreased with lowest value at 4 weeks (all P<0.01). (2)Compared with sham-operation group and normal control group,the relative activation level of Caspase-3 and 12 in CME group increased remarkably (all P<0.05).The time courses of Caspase-3 and 12 expressions corresponded well to those of cardiomyocyte apoptosis after CME. Conclusions The amount of cardiomyocytes apoptosis is significantly increased after CME. Caspase-12 may be involved in the apoptosis of cardiomyocyte after CME.
7.Analysis of infective status of Clonorchis sinensis in Jiangxi Province
Weiming LAN ; Weisheng JIANG ; Kunjiao DAI ; Chunqin HANG ; Shuying XIE ; Jun GE ; Zhaojun LI ; Xiaojun ZENG
Chinese Journal of Schistosomiasis Control 2017;29(1):44-47
Objective To understand the current prevalence of clonorchiasis in Jiangxi Province. Methods A survey was performed according to the scheme of the 3rd Principal Human Parasites of Jiangxi Province. Based on the ecological regions,a stratified cluster sampling method was applied by the economic and geographic situation. In rural areas,the investigation of C. si?nensis was carried out together with the soil?transmitted helminths investigation,and in the urban areas,the random cluster sam?pling method was applied for the C. sinensis investigation. There were 92 survey sites from 32 counties. The eggs of C. sinensis in stool were examined by Kato?Katz technique,and health knowledge was also investigated by questionnaires in some people at the same time. Results A total of 23 606 sample residents were investigated,and 138 were found infected with C. sinensis, with the infection rate of 0.58%. Light infection was found in most of them. Totally 124 C. sinensis infected persons focused in Xinfeng County,and only a few of infected people scattered in the other counties. In Xinfeng County,851 residents were investi?gated. Among them,the infected people were found in all the age groups except the 0?year age group. The highest infection rate appeared in the 70? years group(24.00%). The male infection rate was 20.29%,which was higher than that of the female (6.25%),showing a statistically significant difference(P<0.01). The infection rate was highest in the population who received high school or technical secondary school education(31.48%). For the occupation distribution,the infection rate was highest in public officers(39.39%). The questionnaire survey showed that the infection rate in the populations in Xinfeng County who had the history of eating raw fish or raw shrimp was 33.15%. Conclusions The distribution of C. sinensis infection presents a region?al aggregation in Xinfeng County,but in other areas,the distribution is sporadic. It is necessary to continue to carry out the para?sitic disease screening,and in Xinfeng County,it is necessary to strengthen the comprehensive prevention and control interven?tion.
8.Relationship between endothelial progenitor cells and cardiovascular diseases in maintenance hemodialysis patients
Yaping ZHAN ; Huili DAI ; Weiming ZHANG ; Mingli ZHU ; Yan FANG ; Renhua LU ; Zhaohui NI ; Jiaqi QIAN
Chinese Journal of Nephrology 2016;32(12):881-887
Objective To investigate the relationship between the variation of endothelial progenitor cells (EPC) number and cardiovascular diseases (CVD) in maintenance hemodialysis (MHD) patients ,and discuss the function of EPC in the progression of CVD in MHD. Methods One hundred and fifteen MHD patients over 18 years whose dialysis vintage was over six months from Department of Nephrology, Renji Hospital, Shanghai Jiao Tong University School of Medicine were enrolled. They were divided into CVD group and non ? CVD group by medical history, electrokardiographie (EKG), cardiac ultrasound, peripheral vascular imaging and cardiovascular imaging. Peripheral blood (5 ml) was collected for detecting EPC number by flow cytometry as CD34/CD133/vascular endothelial growth factor receptor 2 (VEGFR2) cells. The EPC number between CVD group and non?CVD group was compared. The relationship between the decrease of EPC number and CVD risks in MHD patients was analyzed by logistic regression analysis. In a three?year follow?up, the death and new CVD events of the two groups were compared in order to discuss the relationship between EPC number and adverse events. Results Among 115 MHD patients, the average age was 61.57 ± 12.76, male/female was 71/44, the average dialysis vintage was (86.24 ± 56.31) months, the average Kt/V was 1.69 ± 0.29 and average ultrafiltration volume was (2.48 ± 0.90) L. Forty?four patients in 115 (38.3%) were with concurrent CVD. The EPC number in CVD group was significantly lower than that in non CVD group (P=0.015). The CVD group had higher serum phosphate (P=0.013), higher glycosylated hemoglobin (P<0.001), but serum calcium, intact parathyroid hormone (iPTH) and other indicators had no significant difference between two groups. Multiple Logistic regression analysis showed that older age (OR=1.061), history of diabetes (OR=9.796), dialysis vintage (OR=1.015), serum phosphate (OR=3.766), decrease of EPC number (OR=0.909) were the independent impact factors of CVD events in MHD patients. There were 22 patients of the 115 MHD patients had encountered a new CVD event in a three?year follow?up between December 2012 and December 2015, 9 patients from the CVD group and 13 patients from the Non?CVD group, and there was no significant difference between two groups (P=0.776). Nine patients from the CVD group and 7 patients from the Non?CVD group died in the follow?up, and there was no significant difference (P=0.111). Seventy?one MHD patients from the non?CVD group were divided into two groups by the median of EPC number. There were 3 patients in the higher EPC number group encountered CVD events and 10 patients in the lower EPC number group encountered CVD events, which had significant difference (P=0.024). Conclusion The decrease of circulating EPC number may be related with CVD events in MHD patients. Even adjusted by age, sex, diabetes, dialysis vintage and serum phosphate, decreased EPC number is still the independent risk factor of CVD events in MHD patients. The decrease of EPC number in MHD patients may be used to predict the occurrence of cardiovascular events.
9.Studies on changes of vegetation and Oncomelania hupensis snails in Poyang Lake after impoundment of Three Gorges Project
Zhaojun LI ; Honggen CHEN ; Xiaojun ZENG ; Yuemin LIU ; Yiyang CHEN ; Kunjiao DAI ; Weiming LAN ; Shuying XIE
Chinese Journal of Schistosomiasis Control 2014;(2):115-120
Objective To explore the impact and change regularity of Oncomelania hupensis snails and vegetation in the marshlands of Poyang Lake caused by the impoundment of the Three Gorges Project. Methods By using the methods of stratified systematic sampling and pinpoint navigation,the succession of vegetation was observed and the distribution of O. hupensis snails was investigated in a period of 8 years on 4 typical marshlands in Poyang Lake. The regularity of changes in vegetation and O. hu-pensis snails was analyzed on the marshlands at high,medium and low elevations,respectively. Results After the impoundment of Three Gorges Project,the average flooding days of these 4 marshlands in 2004 and in 2008 were reduced of 74.8 and 65.8 days compared with those in 2002,the year just before the impoundment of the project,respectively. The change of spatial distribution pattern and the succession of dominant species of vegetation on the marshlands of Poyang Lake have taken place since 2005,and several vegetations showed a trend of moving toward the center of the lake. The hygrophilous vegetation has degenerated and gradu-ally been replaced by mesophily meadow in the marshland area above 14 m elevation. Carex,Phalaris arundinacea and Polygo-num appeared in large area and expanded slowly towards the center of the lake on the marshland below 14 m elevation. O. hupensis snails were mostly found under the Carex association. The tendency of decreasing in density of snails and of migration of snail habi-tats towards low terrain was observed. Conclusions The impoundment of Three Gorges Project has had a significant impact on hy-drological regimes,and a new succession of vegetation has taken place in the marshlands of Poyang Lake. O. hupensis snails and the mesophily vegetation show a trend of migrating slowly to the areas of low elevation in the marshlands.
10.Follow-up of the serum concentration of pituitary hormones after spontaneous subarachnoid hemorrhage
Rongcai LIU ; Qiang HUANG ; Weiming DAI ; Yuanqing JIE ; Guofeng YU ; Xiaofeng FAN ; An WU
Journal of Endocrine Surgery 2014;(6):491-494
Objective To explore the risk of pituitary dysfunction for spontaneous subarachnoid hemor -rhage( s-SAH) patients and to evaluate its correlation with clinical data .Methods 63 cases of patients with s-SAH were selected.Indicators(serum adrenocorti cotrophic hormone (ACTH), growth hormone(GH), insulin-like growth factor-1(IGF-1), thyroid stimulating hormone(TSH), thyroxine(T4), triiodothyronine(T3), folli-cle-stimulating hormone(FSH), luteinizing hormone(LH), total testosterone(in males), estradiol(in females) and prolactin)dynamic observation were tested 7 days, 1 year, 2 years, 3 years after the onset.Results Thirty patients( 47.6%) had some type of pituitary dysfunction .ACTH deficiency was the most frequent disorder (14.3%), followed by GH deficiency(12.7%), hypogonadatrophic(6.3%), hypothyroidism(3.2%)and high prolactin(1.6%).six patients(9.5%)showed deficiencies in more than one axis .Hormone deficiency was relat-ed to vasospasm, Fisher grade 4(P<0.05)and recovery time.Conclusions A greater incidence of hormone de-ficiency after s-SAH was related with vasospasm , Fisher grade and recovery time .Follow-up time should be ex-tended.