1.The induction of tolerogenic dendritic cells by curcumin and its mechanism
Heng-Cheng ZHU ; Xiu-Heng LIU ; Tao TAO ;
Chinese Journal of Organ Transplantation 2005;0(11):-
Objective To observe the induction of tolerogenic dendritic cells (DCs) by curcumin (Cur) and its mechanism.Methods After immature DCs from bone marrow cells of Wistar rats were treated with different concentrations of Cur (0,10,20 and 30?mol/L) respectively,and then the DCs were tested by flow cytometry for the surface molecules expression.After the immature DCs were treated by 30?mol/L Cur with or without stimulation of LPS,endocytosis of DCs to dextran was tested by flow cytometry.The production of IL-12 in DC culture supernatant was determined by ELISA.The levels of NF-?B p65 and RelB translocation to the nucleus were investigated by Western- blot.The activity of NF-?B was detected by NF-?B-binding ELISA and luciferase reporter gene analy- sis.The ability of DCs to stimulate the proliferation of T cells from Lewis rats were analyzed by mixed leukocyte reactions (MLR).Results Cur suppressed LPS-indueed cell-surface expression of costimu- latory molecules (CD80,CD86 and CD40) in a dose-dependent manner.When Cur was used at a con- centration of 30?mol/L,there was no marked difference in the surface molecules expression of LPS- inducing DCs as compared with immature DCs.After DCs were induced by LPS (LPS group),the positive rate of FITC-Dextran uptake was (36.6?7.2)%,and the secretory amounts of IL-12 were (415.9?42.7) pg/ml.In DCs of LPS group,the intranuclear RelB and p65 were highly expressed and their DNA binding activity was 0.65?0.08 and 0.74?0.07 respectively.The luciferase activity of reporter gene in LPS group DCs was remarkably increased to 435% as compared with that in the controls.DCs in LPS group showed strong capacity to stimulate T cells proliferation.When DCs were treated with 30?mol/L Cur followed by induction with LPS (Cur+LPS group),the positive rate of Dextran uptake was (78.6?14.2)% and remarkably higher than in LPS group (P
3.Protective effect of caffeine citrate on neonatal rat brain white matter damage induced by infection
Falin XU ; Heng LI ; Huiqing CHENG ; Xiaojun ZHAO ; Shushuo YANG
Chinese Journal of Applied Clinical Pediatrics 2017;32(14):1085-1088
Objective To investigate the protective effect of caffeine citrate on white matter damage in 2-day-old neonatal rats induced by postnatal infection.Methods Forty-eight 2-day-old SD rats were randomly divided into the control group (group A,n =16),lipopolysaccharide(LPS) infection group (group B,n =16),and caffeine citrate intervention group (group C,n =16) according to the random table method.The newborn rats of group B and C were continuously injected LPS 0.6 mg/kg intraperitoneally for 5 days from 2 days old,and the newborn rats of group A were continuously injected by an equal volume of 9 g/L saline intraperitoneally.Group C was continuously injected by caffeine citrate 10 mg/kg intraperitoneally for 7 days from 4 days old;equal volume of 9 g/L saline was injected into group A and B for 7 days continuously.At 12 days old,8 rats of each group were sacrificed randomly to evaluate the expression of myelin basic protein(MBP) subcortical white matter by immunohistochemical method.Both sides of hippocampus of the rest 8 rats of each group were taken out in ice surface rapidly.The left hippocampus was used to detect the expression of MBP and Caspase-3 by Western blot method,and the right hippocampus was used to evaluate the MBP and Caspase-3 protein level by quantitative real-time PCR(qPCR) method.Results The mean integral optical density (IOD) of subcortical MBP positive expression in group A,group B and group C were 132.64 ± 1.94,102.43 ± 2.12,114.25 ± 2.04,and the difference was statistically significant among 3 groups (F =22.912,P < 0.05).The relative expression levels of MBP mRNA of 3 groups in hippocampus were 0.79 ± 0.01,0.39 ± 0.03,0.55 ± 0.02,and the difference was statistically significant among 3 groups (F =18.584,P < 0.05).The relative expression levels of MBP protein of 3 groups in hippocampus were 0.64 ± 0.03,0.31 ± 0.03,0.51 ± 0.05,and the difference was statistically significant among 3 groups (F =25.780,P < 0.05).The relative expression levels of Caspase-3 mRNA of 3 groups in hippocampus were 0.34 ± 0.02,0.74 ± 0.03,0.57 ± 0.04,and the difference was statistically significant among 3 groups (F =6.105,P < 0.05).The relative expression of Caspase-3 protein of 3 groups in hippocampus were 0.11 ± 0.03,0.36 ± 0.02,0.23 ± 0.03,and the difference was statistically significant among 3 groups (F =40.541,P < 0.05).Conclusions Caffeine citrate has showed protective effect on white matter damage in neonatal rats of 2 days old induced by postnatal infection.The mechanism may be related to inhibition of inflammatory response and reducing apoptosis.
4.Experiment research of natural killer cells amplification in vitro and the killing effect on ovarian cancer cells
Hongyan CHENG ; Xue YE ; Ruiqiong MA ; Xiaohong CHANG ; Heng CUI
Chinese Journal of Obstetrics and Gynecology 2017;52(8):545-550
Objective To amplify natural killer (NK) cells in vitro and explore its killing effect on ovarian cancer cells.Methods (1) The separation of NK cells and identification.A total of 20 ml peripheral blood of one healthy volunteer was collected in Nov.2015,Peking University People's Hospital.The peripheral blood mononuclear cells of normal volunteers were isolated,cultured in vitro and amplificated cultivation for 14 days with K562 cells transfected and expressing interleukin 21 (IL-21-K562) as nourish cells.The number and dynamic state of the growth cells were monitored during the cultured process.Cells were harvested and counted after 14 days cultured.The NK cells phenotypes were detected by flow cytometry.(2) The killing effect of NK cells on ovarian cancer cells:the ratio of effector cells (NK cells) and target cells (ovarian cancer cells and its control) was 50∶ 1,20∶ 1,10∶ 1,5∶1 or 1 ∶ 1,NK cells killing effect on ovarian cancer cells was detected by the lactate dehydrogenase (LDH) release experiments.Results (1) The results of NK cells establishment and phenotypic characterization:the cells were induced in vitro for 14 days by amplification culture.With the extension of incubation time,the number of NK cells increased constantly,from 2.0× 107 on day 0 to 5.1 × 109 on day 14.Obvious amplification of the total number of cells were detected for 255 times.Living cells unstained by trypan blue eventually reached 95% above.Before and after the induction and amplification in vitro,the percentage of NK cells (CD3-CD56+cells) in CD3-cells were 2.33% and 85.32%,respectively (P<0.01),which covered the whole lymphocytes 1.06% and 69.42%,respectively (P<0.01),which showed that NK was the main cell type in the amplificated lymphocytes.(2) The killing rate of NK cells on ovarian cancer cells in vitro:the results detected by LDH release experiments showed that NK cells could performed strong nonspecific killing effect on ovarian cancer cell lines SKOV3,HOC1A,3AO and CAOV3,as well the normal ovarian cell line T29 and NK sensitive cell line K562,and the killing effect increased significantly along with the increase of effector cells and target cells ratio (P<0.01).When the ratio was 1 ∶ 1,the killing rate was 37% for K562,while the rate of killing of other cells was around 10% (P<0.05).When the effect-target ratio was 20∶1 and 50∶ 1,in addition to CAOV3 cells (more than 70%),NK cells had a kill rate of more than 80% for other ovarian cancer cells lines and their control cell K562 and T29 cells (P>0.05).Conclusion NK cells could be established in vitro and have a good non-specific killing effect on ovarian cancer cells.
5.Oropharyngeal Bacterial Carriage in Hospitalized Elderly Patients Without Acute Infections
Heng DONG ; Meifen CHENG ; Jingwen GU ; Minggui WANG
Chinese Journal of Nosocomiology 1994;0(01):-
OBJECTIVE To investigate the oropharyngeal bacterial carriage in hospitalized elderly patients without acute infections and the antimicrobial resistance,and the risk factors for the bacterial carriage.METHODS An oropharyngeal swab was taken from each patient after they rinsed their mouths with sterile saline.Bacteria were cultured and identified with routine methods and the antimicrobial susceptibility testing was carried out with disk diffusion method.RESULTS The oropharyngeal bacterial carriage rate was 55.2% in elderly patients.Sixty two pathogens were isolated including 56(90.3%) strains of Gram-negative bacilli,5(8.1%) Gram-positive cocci and 1 Candida albicans.All of 34 strains of Haemophilus spp were susceptible to antimicrobials tested such as ampicillin and cefaclor,a few strains were resistant to gentamicin and ciprofloxacin.Of 10 strains of Klebsiella pneumoniae,4 strains were extended-spectrum ?-lactamases positive and resistant to cefotaxime.Logistic analysis indicated that denture-wearing was the risk factor for the oropharyngeal bacterial carriage in elderly.CONCLUSIONS Oropharyngeal bacterial carriage rate is high in hospitalized elderly patients.The major colonized bacteria are Gram-negative bacilli.Denture-wearing is the risk factor for the oropharyngeal bacterial carriage in elderly.
6.Synthesis and biological evaluation of H2S donor ADT-OH derivatives
Yuyao LI ; Heng SONG ; Jian CHENG ; Guizhen AO
Journal of China Pharmaceutical University 2017;48(3):276-281
5-(4-Hydroxyphenyl)-3H-1,2-dithiole-3-thione (ADT-OH) is a slowly releasing H2S donor with some neuroprotective effect.In order to study the structure-activity relationships,seventeen compounds (Y1-Y17) were synthesized by modification of ADT-OH at the aromatic ring position,and their structures were confirmed by 1H NMR,13C NMR and HR-MS.Among them,6 compounds (Y4,Y13-Y17) were novel compounds.Their effects had been evaluated on HT-22 hippocampal neuron cells damaged by glutamate with MTF method.The pharmacological results demonstrated that all the Y compounds had potent neuroprotection at most of the tested concentrations (1-100 μmol/L).Compounds Y1-Y9 and Y11 significantly improved the survival rates of the damaged cells at 1-100 μmol/L (P <0.01).Specially,compounds Y1,Y4,Y6-Y9,Yu are more potent than their parent compound ADT-OH at concentration of 1-10 μmol/L,which is worthy of further study.
7.Considerations on the 5-in-1 social co-governance for drug safety
Yunwu ZHAO ; Heng WANG ; Jingya WU ; Cheng BIAN ; Niannian LI
Chinese Journal of Hospital Administration 2015;31(1):39-41
In this consideration,the paper described the imperatives of drug safety,constraints of governance by the government,and advantages of social co-governance,proposing the necessity to establish aFive-in-One (referring to the involvement of the government,enterprises,industry associations,media and the public) drug safety social co-governance.In its analysis of the present dilemma o[drug safety governance,the authors proposed such measures as perfecting legal and institutional system,reforming governmental power allocation mechanism,improving the incentive mechanism,innovating governance means and improving social governance ability,for the purpose of enhancing such a co-governance pattern.
8.Application of flexible ureteroscopy combined with Holmium laser lithotripsy for kidney calculi patients with HIV positive
Chuanyu CHENG ; Heng HE ; Erjun ZHANG ; Yudong WU ; Bingqian LIU
Chinese Journal of Urology 2016;37(12):928-931
Objective To discuss the efficacy of the flexible ureteroscopy combined with Holmium laser lithotripsy for kidney calculi with HIV positive patients.Methods From May 2015 to May 2016,47 cases of patients with renal calculi were treated by the flexible ureteroscopy combined with Holmium laser lithotripsy in our hospital.There were 29 cases male,18 cases female,aged from 22 to 56 years old,average 39 years.There were 38 cases with single stone,9 cases with multiple stones.There were 9 cases with stones on both sides.Flexible ureteroscopy channel sheath was used in surgery.Stones were fragmented by Holmium laser.During one-month follow-up after surgery,stone removal and stone free rate were recorded.Residual stones were re-treated with a secondary lithotripsy or ESWL.Results The flexible ureteroscopy channel sheath was indwelled successfully in all the cases.All stones were detected.The average operation time was 63min (range,42-141min) and the median postoperative stay was 4.5days (range,2-16 d).Among the 47 patients,41 patients underwent first-stage lithotripsy,6 patients underwent second-stage ESWL after lithotripsy,and 1 patient underwent third-stage lithotripsy.The one-month stone free rate was 87.2% (41/47).The total stone free rate was 97.9% (46/47) after second-stage lithotripsy.Postoperative fever occurred in 4 cases after lithotripsy.No blood transfusion,systemic infection,ureteral perforation,or ureteral avulsion occurred.The total complication rate was 8.5% (4/47).The mean number of CD4 +T lymphocytes before lithotripsy was 402/μl,and was 410/μl 3 days after lithotripsy.There was no Statistical differences between them.Conclusions Flexible ureteroscopy combined Holmium laserlithotripsy could be a safe and effective treatment for kidney calculi patients with HIV/AIDS positive.
9.Effect of low concentration of cerium oxide nanomaterials on hepatocellular carcinoma cell proliferation
Heng CHENG ; Zhongli LIAO ; Hongyan CHEN ; Tao WANG ; Hong GUO
Journal of Regional Anatomy and Operative Surgery 2016;25(4):244-247
Objective To study the impact of different low concentrations of cerium oxide for hepatocellular carcinoma cell prolifera-tion.Methods Three different types of hepatoma cells (Huh7, HepG2,7721) were cultured,and added different concentrations of cerium oxide (0.005,0.01,0.05,0.1,1 μg/mL),of which the cell proliferation was detected by CCK8.The apoptosis-related genes was detected by qRT-PCR technology.The cell cycle was analyzed by flow cytometry.And the effect of low concentration cerium oxide on hepatocellular carci-noma cells tumorigenicity was confirmed by the nude mice experiments.Results CCK8 experiment showed that low concentrations of cerium oxide could promote proliferation of hepatocellular carcinoma cell, especially in concentration of 0.01μg/mL.The qRT-PCR showed that low concentration of cerium oxide could inhibit the apoptosis of hepatocellular carcinoma cell.The flow cytometry analysis had not found any effect of cerium oxide on cell cycle.The tumorigenicity experiments confirmed that low concentrations of cerium oxide could enhance the tumorigenic ability of hepatocellular carcinoma cell.Conclusion Low concentration of cerium oxide can significantly improve the proliferation of liver cancer cells.
10.Risk factors for hydrocephalus after moderate to severe traumatic brain injury
Cheng CAO ; Jiqiang MA ; Yumin LIANG ; Wei WU ; Heng GAO
Chinese Journal of Trauma 2016;32(7):602-606
Objective To investigate the risk factors of posttraumatic hydrocephalus (PTH) in patients with moderate to severe traumatic brain injury (TBI).Methods Aretrospective study was conducted for 183 patients with moderate to severe TBI (125 males,58 females;6-91 years of age,mean 48.23 years).According the presence of PTH,the patients were allocated into PTH group (n =34) and non-PTH group (n =149).Risk factors of PTh were assessed by univariate and logistic regression analysis,including gender,age,injury types,injury severity,intraventricular hemorrhage,subarachnoid hemorrhage,midline shift,subdural effusion,therapeutic strategies and skull defect.Association between the boundaries of skull defect and PTH was determined.Results Between-group differences were not significant regarding age,gender,injury types and intraventricular hemorrhage (P > 0.05),but differed significantly in injury severity,subarachnoid hemorrhage,midline shift,subdural effusion,craniectomy and skull defect (P < 0.05).Further Logistic regression analysis confirmed subarachnoid hemorrhage (OR =6.169),interhemispheric subdural effusion (OR =31.743),and unilateral (OR =17.602) and bilateral (OR =30.567) skull defects were risk factors of PTH.Of the patients with unilateral skull defect following decompressive craniectomy,the inferior limit ≤ 10 mm from the zygomatic arch also played a role in the development of PTH (OR =5.500,P < 0.05).Conclusions Subarachnoid hemorrhage,interhemispheric subdural effusion and skull defect are risk factors of PTH.Unilateral skull defects with the inferior limit too close to the zygomatic arch can predispose to the development of PTH.