1.Differentiation of bone marrow mesenchymal stem cells into endometrial epithelial cells in vitro
Chinese Journal of Tissue Engineering Research 2016;20(19):2770-2776
BACKGROUND:Previous studies have confirmed that as an exogenous origin of endometrial epithelial cel s, bone marrow stem cel s are involved in the functional reconstruction of the injured endometrium. OBJECTIVE:To discuss whether bone marrow mesenchymal stem cel s can differentiate into endometrial epithelial cel s in vitro. METHODS:Passage 2 bone marrow mesenchymal stem cel s and endometrial stromal cel s were col ected and subjected to different treatments:bone marrow mesenchymal stem cel s were cultured alone in the DMEM/F12 medium containing 2%fetal bovine serum or in the DMEM/F12 medium containing 2%fetal bovine serum, 10-7 mol/Lβ-estradiol and 10μg/L epidermal growth factor as groups 1 and 2, respectively;bone marrow mesenchymal stem cel s co-cultured with endometrial stromal cel s by Transwel chamber were cultured in the DMEM/F12 medium containing 2%fetal bovine serum or in the DMEM/F12 medium containing 2%fetal bovine serum, 10-7 mol/Lβ-estradiol and 10μg/L epidermal growth factor as groups 3 and 4, respectively. After 5 days culture, bone marrow mesenchymal stem cel s at the bottom of the culture plate were col ected to detect expressions of epithelial cel markers, such as CK7, CK18, CK1 and EMA by RT-PCR technology, and to determine keratin expression using immunofluorescence assay. Besides, expressions of these epithelial cel markers in the group 3 were detected using RT-PCR at 1, 3, 5 and 7 days of culture, respectively. RESULTS AND CONCLUSION:mRNA expression of CK7 showed a significant successive rise in the groups 1, 2, 3, 4 and it was highest in the group 4. mRNA expressions of CK18 and CKl9 in the later three groups had no significant differences, but al significantly higher than those in the group 1. Compared with the groups 1 and 3, mRNA expression of EMA in the groups 2, 4 were significantly higher, but no significant differences existed between groups. Expression of keratin was strongly positive in the group 4, weakly positive in the group 3 and negative in the groups 1, 2, respectively. Furthermore, with the increase of culture time, mRNA expression of CK7 exhibited a constant increase, which was significantly higher at 5 and 7 days than at 1 and 3 days;but there were no significant differences in expressions of CKl9, CKl8 and EMA at different time points. These results show that bone marrow mesenchymal stem cel s can differentiate into endometrial epithelial cel s in vitro under certain conditions. Moreover, it can remarkably promote the endometrial differentiation under the combined effects of some exogenous and endogenous factors.
2.Experimental Study on Bortezomib for Treatment of Ulcerative Colitis
Chinese Journal of Gastroenterology 2017;22(7):402-406
Background:The efficacy of traditional medicine on ulcerative colitis (UC) is often unsatisfactory, hence development of drug based on the pathogenic mechanism of UC becomes a hot topic in the research of UC.It has been revealed in recent studies that activation of nuclear factor-κB (NF-κB) is implicated as a key regulator in the immune and inflammatory responses in UC.Aims:To explore whether bortezomib, a potent proteasome inhibitor that inhibits NF-κB activation can be used for treatment of UC.Methods:Thirty-two BALB/c mice were used to induce acute experimental colitis by drinking 3%dextran sulfate sodium (DSS) freely for 7 days, and then randomly allocated into four groups injected intraperitoneally with 0.2 (low-dose group), 0.6 (medium-dose group), 1.0 mg/kg (high-dose group) bortezomib and normal saline (model control group), respectively.On the 7th day after treatment, the disease activity index (DAI) and histopathological change of colonic tissue were observed;the colitis-related parameters including peripheral blood hemoglobin (Hb), C-reactive protein (CRP) and colonic myeloperoxidase (MPO) activity were measured, and the nuclear translocation of NF-κB was assessed by electrophoretic mobility shift assay.Results:Compared with the model control group, the DAI, CRP, MPO activity, and injury score of colonic tissue were decreased gradually, and the Hb was increased gradually in mice treated with low-, medium-and high-dose bortezomib (P all <0.05).The efficacy of medium-and high-dose bortezomib was notable.In mice treated with medium-and high-dose bortezomib, nuclear translocation of NF-κB was inhibited obviously.Conclusions:Bortezomib can modulate the colonic inflammation in mice with experimental colitis by inhibiting NF-κB activation and subsequently improving the clinical manifestations, colitis-related parameters and tissue damage.Increasing the dosage of bortezomib in a safety range may enhance the treatment response.
3.The study on the level of serum selenium with infantile diarrhea
Hui QU ; Kai KANG ; Shuli FU ; Lihong REN
Chinese Journal of Postgraduates of Medicine 2012;35(30):12-14
Objective To explore the relationship of the level of serum selenium and infantile diarrhea,provide foundation for establishing the therapeutic criteria.Methods Seventy-eight diarrhea children was enrolled in this study,6 -24 months old.Thirty children with acute diarrhea (AD group),26 children with persistent diarrhea (PD group) and 22 children with chronic diarrhea (CD group).The level of serum selenium was measured and compared with another 30 healthy children (control group) of matched sex and age.The level of serum selenium of CD group was compared before and after the recovery.Results The level of serum selenium in AD group and PD group had no significant difference compared with control group [ (51.34 ± 4.84),(48.14 ± 3.05 ) μ g/L vs.(55.08 ± 5.59 ) μ g/L ] (P>0.05 ).But the level of serum selenium in CD group was significantly lower than that in control group [ (42.13 ± 5.16) μ g/L vs.(55.08 ±5.59) μg/L] (P<0.05).After treatment for 1 month,the level of serum selenium in CD group was significantly increased than before treatment [ (53.76 ± 8.38 ) μ g/L vs.(42.13 ± 5.16) μ g/L ] (P<0.05 ).Conclusions The nosogenesis of chronic diarrhea may relate with the level of serum selenium decrease.Therapeutic selenium supplement is important in children with chronic diarrhea.
4.Relationship between expression of integrin ?1,?1 and apoptosis in placenta of pregnancy induced hypertension syndrome
Xia CAI ; Guangdi QU ; Lihong GENG ; Al ET
Chinese Journal of Perinatal Medicine 1998;0(01):-
Objective To investigate the expression of integrin ?1 and ?1 and to study their role on cell apoptosis of placenta with pregnancy induced hypertension syndrome. Methods Immuno histochemistry was used for 40 placental samples with PIH and 43 normal placental samples to detect the expression of integrin ?1、?1; TDT mediated dUTP nick end labeling(Tune1) was used for 12 placental samples with PIH and 24 normal placental samples to examine apoptosis index in cytotrophoblast, syncytiotrophoblast and decidual cell. Results The apoptosis index in placental cytotrophoblast, syncytiotrophoblast and decidual cells in PIH group were (11.04 ? 3.46)%, (12.2 ? 3.67)%, (13.03 ? 4.38)% respectively and was significantly higher than of control group [(3.91 ? 1.65)%,(5.39 ? 1.76)%, (4.08 ? 1.97)%] ( P
5.Porhyromonas endodontalis liopolysaccharide regulates interleukin-23 expression in mouse osteoblasts
Liu QU ; Yaqiong YU ; Lihong QIU ; Nan MA ; Ming ZHONG
Journal of Practical Stomatology 2016;32(4):453-456
Objective:To investigate the effects of Porhyromonas endodontalis(P.e)liopolysaccharide(LPS)on the expression of IL-23mRNA and protein in mouse osteoblasts MC3T3-E1 and the role of phosphatidylinositol 3-Kinases (PI3K)signaling pathway in this process.Methods:MC3T3-E1 cells were treated with different concentrations of P.e LPS for different hours,or pretreated with LY294002,a special PI3K inhibitor.The IL-23 mRNA and protein expression levels were detected by real-time PCR and Western blot respectively.Results:The level of IL-23 mRNA increased in MC3T3-E1 cells with the increase of concentration and the treatment time of P.e LPS(P <0.05).The IL-23 protein expression was increased by P.e LPS in a time dependent manner(P <0.05).The mRNA and protein of IL-23 decreased(P <0.05)after pretreatment with LY294002.Conclusion:P.e LPS can induce the expression of IL-23 mRNA and protein in MC3T3-E1 cells,and the PI3K signaling pathway may play a part in this process.
6.Establishment of the reference interval for CRE, UREA among healthy college students
Bing ZHAO ; Jirong LIU ; Jing SUN ; Lihong YANG ; Feifei QU
Chinese Journal of Primary Medicine and Pharmacy 2016;23(6):848-851
Objective To analyze the CRE and UREA result of university students,and establish the refer-ence interval for CRE,UREA adapted to this university.Methods Performing serum analysis of 2 926 freshmen for CRE,UREA with the automatic biochemical analyzer ( Olympus AU400 ) .Results Reference intervals were get among freshmen as CRE:74.00-109.00μmol/L (M),59.00-85.00 μmol/L (F),and it was different between male and female(Z =-27.27,P <0.01);UREA:2.70 -6.70mmol/L (male aged 16 -24 years),1.90 -5.70mmol/L (female aged 16-24 years),2.30-6.57mmol/L (male aged >24-45 years),1.94-5.11mmol/L ( female aged 25-45 years) .Conclusion The reference intervals disaggregated by sex and age are initially estab-lished.
7.Application of activated T lymphocytes induced by hepatocellular carcinoma rejection antigenic peptide-pulsed DCs in vivo and in vitro
Ping QU ; Yunxin CAO ; Yanfang SUI ; Lihong ZHANG ; Xiumin ZHANG ;
Chinese Pharmacological Bulletin 1986;0(04):-
AIM To study if the activated T lymphocytes induced by hepatocellular carcinoma (HCC) rejection antigenic peptide SLIVHLNEV (mutant peptide of C met) pulsed dendritic cells (DCs) can provide protection against tumor challenge in nude mice HCC model and in vitro . METHODS SLIVHLNEV was eluted from HCC cell line HHCC by mild acid buffer and was identified by the mass spectrometry technique. SLIVHLNEV pulsed DCs isolated from HLA A2+ ruptured spleen were co cutured with isogeneic T lymphocytes. Cytotoxicity was tested using lactate dehydrogenase (LDH) release assay. The induced cytotoxic T lymphocytes (CTLs) were implanted into nude mice in order to protect them against transplanted HHCC tumor cells. The inhibition effect of CTLs on the growth of implanted tumor in nude mice was also observed. RESULTS The CTLs induced by SLIVHLNEV pulsed DCs had unique ability to kill HHCC tumor cells in vitro. They also protected the nude mice against the further challenge of HHCC tumor cells and inhibited the growth of implanted tumor. CONCLUSION DC based HCC rejection antigenic peptide SLIVHLNEV vaccine can induce powerful immune reaction both in nude mice HCC model and in vitro.
8.Phase Ⅰ study of weekly PTX+DDP, and postoperative radiotherapy for early cervical cancer in Chinese women
Lihong ZHU ; Suqing TIAN ; Ang QU ; Hao WANG ; Junjie WANG ; Hongyan GUO
Chinese Journal of Radiation Oncology 2016;25(8):834-838
Objective To determine the maximum tolerated dose ( MTD) and dose?limiting toxicity ( DLT) of weekly PTX and DDP concurrent postoperative radiotherapy in Chinese women with high?and intermediate?risk early cervical cancer. Methods Women with high risks postoperative cervical carcinoma, ECOG≤2 were eligible. Pelvis RT (6/10 MV X?ray,3DCRT 40 Gy/20f,para?metrial boost 10?20 Gy/5?10f) was followed by 2?4f brachytherapy applications ( 192 Ir,5 Gy/f) . Concurrent weekly chemotherapy was started at DDP 20 mg/m2 and PTX 10 mg/m2 weekly,and escalated in three?patient cohorts according to 3+3 methods. Results 25 patients were enrolled and treated over seven doses levels until dose?limiting toxicity (DLT) was reached. Median age was 48 years (range,34?66).All of patients finished RT in 7 weeks. Grade 3,4 non?hematologic toxicities were diarrhea and observed in two patients (4 cycles,DLT) at level 7.Grade 3,4 hematologic,principally leukopenia and neutropenia,and occurs late cycles. One grade 4 leukopenia and neutropenia was observed at dose level 6 but not seen in three additional patients. No one was delayed treatment time by concurrent chemotherapy.22 patients finished 6 cycles. Median follow?up is 59. 5 months. Three patients have died of cancer metastasis and recurrence. One patient has died of respiratory failure. Conclusions Combination PTX and DDP administered concurrently with pelvic EBRT can be safely administered at the MTD of DDP 35 mg/m2 and PTX 30 mg/m2 weekly for six cycles in Chinese women with postoperative cervical cancer.
9.Influence of two enteral nutrition injection methods on prognosis of acute cerebral infarction induced pseudobulbar palsy
Wenxiang XU ; Xifang ZHU ; Xiuhua WU ; Shuqin LIANG ; Lihong QU ; Jing SUN
Chinese Journal of Practical Nursing 2016;32(20):1525-1530
Objective To investigate two enteral nutrition injection methods on prognosis of patients with acute cerebral infarction (ACI) induced pseudobulbar palsy patients. Methods A total of 86 patients of ACI accompanied with dysphagia were randomly divided into the feeding pump group (n=43) and the traditional nasal feeding group (n=43). Nutritional support for both groups lasted at least more than 14 d through the feeding tube. In the first and fourteenth day data of both groups concerned with body mass index (BMI), triceps skin fold (TSF), upper arm muscle circumference of triceps (MAMC), heamoglobin (Hb), serum prealbumin (Alb), alanine transaminase(ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), creatinine (Cr), incidence of complications, neuro-function score NIHSS and the swallowing function evaluation were collected and analyzed. Results Nutritional index Hb and Alb was (132.6±10.1) g/L and (38.2±1.4) g/L in the group of the feeding pump group, and significantly higher than (122.2±13.2) g/L and (36.3±1.5) g/L in the traditional nasal feeding group in the fourteenth day, the difference was significant (t=12.86, 12.37, P<0.05). The NHISS score was 5.2±1.0 in the feeding pump group, significantly lower than 8.9±1.1 of the traditional nasal feeding group, the difference was significant (t=-26.37, P < 0.05). The improved rate of swallowing function was 81.4% (35/43) in the feeding pump group, significantly higher than 60.5%(26/43) of the traditional nasal feeding group, the difference was significant (χ2=4.568, P < 0.05). Conclusions The use of enteral feeding pump at early stage for ACI patients with pseudobulbar palsy can improve the nutritional status, acute neurological function, swallowing function and lower the incidence of complications which is beneficial for early recovery and short-term prognosis.
10.Therapeutic effects of human umbilical cord mesenchymal stem cell transplantation on acute liver failure in rats
Hongying GUO ; Hui ZHU ; Zhengguo ZHANG ; Lihong QU ; Hong XIAO ; Yanling FENG ; Ming XIANG ; Fangming LI ; Xiang HU ; Jiefei WANG
Chinese Journal of Infectious Diseases 2011;29(5):261-266
Objective To study the therapeutic effects of human umbilical cord mesenchymal stem cells (hUCMSCs) on acute liver failure ( ALF) induced by D-galactosamine (D-gal) and lipopolysaccharide (LPS) in rats. Methods The ALF model was obtained through intraperitoneal injection of D-gal(300 mg/kg)and LPS (20μg/kg)in Wister rats. The hUCMSCs were transplanted after intoxication. All rats were divided into four groups, and each group received either hUCMSCs or 0.9% NaCl solution through intraperitoneal or tail-intravenous injection. To evaluate the liver function of each group, the levels of alanine aminotransferase (ALT), total bilirubin (TBil) and serum albumin (Alb) were measured on the day of hUCMSCs transplantation and the following 1, 2, 3, 5 and 7 days. All rats were then sacrificed to examine the liver histology at day 7. Analyses were done by using Fisher's exact test, unpaired t test and Mann-Whitney U test. Results There were no significant differences of survival rates among four groups (Fisher's exact test, both P = 1. 00). The levels of ALT, TBil and Alb in group receiving hUCMSCs intraperitoneally were (804. 9 ± 88. 0) U/L,(17. 4±2. 7) μmol/L and (20. 9±0. 8) g/L, respectively after 2 days of injection, whereas in the corresponding control group, those were (1294. 3± 171. 4) U/L, (32. 3±5. 5) μmol/L and (16. 1±0. 9) g/L, respectively, which indicated that hUCMSCs transplantation significantly improved the liver function (t = 2. 640, P =0.020;t=2.529, P = 0. 025;t= - 3. 833, P = 0. 002). Both of hUCMSCs-transplanted groups showed no significant differences. Liver histological data showed that transplantation of hUSMSCs through either intraperitoneal or tail-intravenous injection alleviated liver damage (U=4. 500, P = 0. 005;U=4. 500, P = 0. 008) and the mitotic index also increased in hUCMSCs-transplanted groups (U=4. 000, P = 0. 005; U=5. 500, P = 0. 013). Conclusions The levels of ALT, TBil and Alb can rapidly normalize in ALF rats after injected with hUCMSCs either intraperitoneally or tail-intravenously. hUCMSCs application raises the mitotic index, enhances hepatocellular regeneration and improves histological status.